Glycopeptide phosphonate derivatives

ABSTRACT

Disclosed are glycopeptides that are substituted with one or more substituents each comprising one or more phosphono groups; and pharmaceutical compositions containing such glycopeptide derivatives. The disclosed glycopeptide derivatives are useful as antibacterial agents.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.11/584,908, filed Oct. 23, 2006; now U.S. Pat. No. 7,351,691 whichapplication is a continuation of U.S. application Ser. No. 11/265,570,filed on 2 Nov. 2005 (now U.S. Pat. No. 7,208,471 B2); which applicationis a continuation of U.S. application Ser. No. 11/037,934, filed 18 Jan.2005 (now U.S. Pat. No. 7,008,923 B2); which application is acontinuation of U.S. application Ser. No. 10/669,778, filed 24 Sep.2003, (now U.S. Pat. No. 6,887,976 B2); which application is acontinuation of U.S. application Ser. No. 09/847,042, filed 1 May 2001(now U.S. Pat. No. 6,635,618 B2); which application claims the benefitof U.S. Provisional Application Ser. No. 60/213,410, filed 22 Jun. 2000;which applications are incorporated herein by reference in theirentirety.

BACKGROUND OF THE INVENTION Field of the Invention

This invention is directed to novel phosphonate derivatives ofglycopeptide antibiotics and related compounds. This invention is alsodirected to pharmaceutical compositions containing such glycopeptidephosphonate derivatives, methods of using such glycopeptide phosphonatederivatives as antibacterial agents, and processes and intermediatesuseful for preparing such glycopeptide phosphonate derivatives.

BACKGROUND

Glycopeptides (e.g. dalbaheptides) are a well-known class of antibioticsproduced by various microorganisms (see Glycopeptide Antibiotics, editedby R. Nagarajan, Marcel Dekker, Inc. New York (1994)). These complexmulti-ring peptide compounds are very effective antibacterial agentsagainst a majority of Gram-positive bacteria. Although potentantibacterial agents, the glycopeptides antibiotics are not used in thetreatment of bacterial diseases as often as other classes ofantibiotics, such as the semi-synthetic penicillins, cephalosporins andlincomycins, due to concerns regarding toxicity.

In recent years, however, bacterial resistance to many of thecommonly-used antibiotics has developed (see J. E. Geraci et al., MayoClin. Proc. 1983, 58, 88-91; and M. Foldes, J. Antimicrob. Chemother.1983, 11, 21-26). Since glycopeptide antibiotics are often effectiveagainst these resistant strains of bacteria, glycopeptides such asvancomycin have become the drugs of last resort for treating infectionscaused by these organisms. Recently, however, resistance to vancomycinhas appeared in various microorganisms, such as vancomycin-resistantenterococci (VRE), leading to increasing concerns about the ability toeffectively treat bacterial infections in the future (see HospitalInfection Control Practices Advisory Committee, Infection ControlHospital Epidemiology, 1995, 17, 364-369; A. P. Johnson et al., ClinicalMicrobiology Rev., 1990, 3, 280-291; G. M. Eliopoulos, European J.Clinical Microbiol., Infection Disease, 1993, 12, 409-412; and P.Courvalin, Antimicrob. Agents Chemother, 1990, 34, 2291-2296).

A number of derivatives of vancomycin and other glycopeptides are knownin the art. For example, see U.S. Pat. Nos. 4,639,433; 4,643,987;4,497,802; 4,698,327; 5,591,714; 5,840,684; and 5,843,889. Otherderivatives are disclosed in EP 0 802 199; EP 0 801 075; EP 0 667 353,WO 97/28812; WO 97/38702; WO 98/52589; WO 98/52592; and in J. Amer.Chem. Soc., 1996, 118, 13107-13108; J. Amer. Chem. Soc., 1997, 119,12041-12047; and J. Amer. Chem. Soc., 1994, 116, 4573-4590.

Despite the above referenced disclosures, a need currently exists fornovel glycopeptide derivatives having effective antibacterial activityand an improved mammalian safety profile. In particular, a need existsfor glycopeptide derivatives which are effective against a wide spectrumof pathogenic microorganism, including vancomycin-resistantmicroorganisms, and which have reduced tissue accumulation and/ornephrotoxicity.

SUMMARY OF THE INVENTION

The present invention provides novel glycopeptide phosphonatederivatives having highly effective antibacterial activity and animproved mammalian safety profile. More specifically, the glycopeptidephosphonate derivatives of the invention unexpectedly exhibit reducedtissue accumulation and/or nephrotoxicity when administered to a mammal.

Accordingly, this invention provides glycopeptide compounds substitutedwith one or more (e.g., 1, 2 or 3) substituents comprising one or more(e.g., 1, 2 or 3) phosphono (—PO₃H₂) groups; or a pharmaceuticallyacceptable salt, stereoisomer, or prodrug thereof. Preferably, theglycopeptide compound is substituted with one or two substituentscomprising one or two phosphono groups. More preferably, theglycopeptide compound is substituted with one substituent comprising oneor two phosphono groups, preferably one phosphono group. Optionally, theglycopeptide compounds of this invention may also be substituted withother substituents not comprising a phosphono group, provided that atleast one substituent comprises one or more phosphono groups.

Accordingly, in one preferred embodiment, this invention provides aglycopeptide compound substituted at the C-terminus with a substituentcomprising one or two phosphono (—PO₃H₂) groups; or a pharmaceuticallyacceptable salt, stereoisomer, or prodrug thereof. Preferably, thephosphono-containing substituent is attached to the carbonyl group atthe C-terminus through an amide bond, an ester bond, or a thioesterbond; more preferably, through an amide bond. Preferably, thephosphono-containing substituent comprises one phosphono group.Particularly preferred phosphono-containing substituents at theC-terminus include phosphonomethylamino, 3-phosphonopropylamino and2-hydroxy-2-phosphonoethylamino.

In another preferred embodiment, this invention provides a glycopeptidecompound substituted at the R-terminus (on the resorcinol ring) with asubstituent comprising one or two phosphono (—PO₃H₂) groups; or apharmaceutically acceptable salt, stereoisomer, or prodrug thereof.Preferably, the phosphono-containing substituent is attached to theR-terminus (i.e., the resorcinol ring) through the nitrogen atom of anaminomethyl group attached to the R-terminus. Preferably, thephosphono-containing substituent comprises one phosphono group.Particularly preferred phosphono-containing substituents at theR-terminus include N-(phosphonomethyl)aminomethyl;-2-hydroxy-2-phosphonoethyl)aminomethyl;N-carboxymethyl-N-(phosphonomethyl)aminomethyl;N,N-bis(phosphonomethyl)aminomethyl; andN-(3-phosphonopropyl)aminomethyl.

In still another preferred embodiment, this invention provides aglycopeptide compound substituted at the C-terminus and at theR-terminus with a substituent comprising one or two phosphono (—PO₃H)groups; or a pharmaceutically acceptable salt, stereoisomer, or prodrugthereof. Preferably, the phosphono-containing substituents eachcomprises one phosphono group.

A preferred compound of the invention is a glycopeptide of formula I:

wherein:

R¹ is selected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl, heterocyclic and —R^(a)—Y—R^(b)—(Z)_(x);or R¹ is a saccharide group optionally substituted with—R^(a)—Y—R^(b)—(Z)_(x), R¹, —C(O)R¹, or —C(O)—R^(a)—Y—R^(b)—(Z)_(x);

R² is hydrogen or a saccharide group optionally substituted with—R^(a)—Y—R^(b)—(Z)_(x), R^(f), —C(O)R^(f), or—C(O)—R^(a)—Y—R^(b)—(Z)_(x);

R³ is —OR^(c), —NR^(c)R^(c), —O—R^(a)—Y—R^(b)—(Z)_(x),—NR^(c)—R^(a)—Y—R^(b)—(Z)_(x), —NR^(c)R^(c), or —O—R^(c); or R³ is anitrogen-linked, oxygen-linked, or sulfur-linked substituent thatcomprises one or more phosphono groups;

R⁴ is selected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,—R^(a)—Y—R^(b)—(Z)_(x), —C(O)R^(d) and a saccharide group optionallysubstituted with —R^(a)—Y—R^(b)—(Z)_(x), R^(b), —C(O)R^(f), or—C(O)—R^(a)—Y—R^(b)—(Z)_(x), or R⁴ and R⁵ can be joined, together withthe atoms to which they are attached, form a heterocyclic ringoptionally substituted with —NR^(c)—R^(a)—Y—R^(b)—(Z)_(x);

R⁵ is selected from the group consisting of hydrogen, halo,—CH(R^(c))—NR^(c)R^(c), —CH(R^(c))NR^(c)R^(c),—CH(R^(c))—NR^(c)—R^(a)—Y—R^(b)—(Z)_(x), —CH(R^(c))—R^(x),—CH(R^(c))—NR^(c)—R^(a)—C(═O)—R^(x), and a substituent that comprisesone or more phosphono groups;

R⁶ is selected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,—R^(a)—Y—R^(b)—(Z)_(x), —C(O)R^(d) and a saccharide group optionallysubstituted with —R^(a)—Y—R^(b)—(Z)_(x), R^(f), —C(O)R^(f), or—C(O)—R^(a)—Y—R^(b) (Z)_(x), or R⁵ and R⁶ can be joined, together withthe atoms to which they are attached, form a heterocyclic ringoptionally substituted with —NR^(c)—R^(a)—Y—R^(b)—(Z)_(x);

R⁷ is selected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,—R^(a)—Y—R^(b)—(Z)_(x), and —C(O)R^(d);

R⁸ is selected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic;

R⁹ is selected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic;

R¹⁰ is selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic; or R⁸ and R¹⁰ arejoined to form —Ar¹—O—Ar—, where Ar¹ and Ar² are independently aryleneor heteroarylene;

R¹¹ is selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic, or R¹⁰ and R¹¹ arejoined, together with the carbon and nitrogen atoms to which they areattached, to form a heterocyclic ring;

R¹² is selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl, heterocyclic, —C(O)R^(d), —C(NH)R^(d),—C(O)NR^(c)R^(c), —C(O)OR^(d), —C(NH)NR^(c)R^(c),—R^(a)—Y—R^(b)—(Z)_(x), and —C(O)—R^(a)—Y—R^(b)—(Z)_(x), or R¹¹ and R¹²are joined, together with the nitrogen atom to which they are attached,to form a heterocyclic ring;

R¹³ is selected from the group consisting of hydrogen or —OR¹⁴;

R¹⁴ is selected from hydrogen, —C(O)R^(d) and a saccharide group;

each R^(a) is independently selected from the group consisting ofalkylene, substituted alkylene, alkenylene, substituted alkenylene,alkynylene and substituted alkynylene;

each R^(b) is independently selected from the group consisting of acovalent bond, alkylene, substituted alkylene, alkenylene, substitutedalkenylene, alkynylene and substituted alkynylene, provided R^(b) is nota covalent bond when Z is hydrogen;

each R^(c) is independently selected from the group consisting ofhydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl,alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl, heterocyclicand —C(O)R^(d);

each R^(d) is independently selected from the group consisting of alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic;

R^(e) is a saccharide group;

each R^(f) is independently alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,heteroaryl, or heterocyclic;

R^(x) is an N-linked amino saccharide or an N-linked heterocycle;

X¹, X² and X³ are independently selected from hydrogen or chloro;

each Y is independently selected from the group consisting of oxygen,sulfur, —S—S—, —NR^(c)—, —S(O)—, —SO₂—, —NR^(c)C(O)—, —OSO₂—, —OC(O)—,—NR^(c)SO₂—, —C(O)NR^(c)—, —C(O)O—, —SO₂NR^(c)—, —SO₂O—,—P(O)(OR^(c))O—, —P(O)(OR^(c))NR^(c)—, —OP(O)(OR^(c))O—,—OP(O)(OR)NR^(c)—, —OC(O)O—, —NR^(c)C(O)O—, —NR^(c)C(O)NR^(c)—,—OC(O)NR^(c)—, —C(═O)—, and —NR^(c)SO₂NR^(c)—;

each Z is independently selected from hydrogen, aryl, cycloalkyl,cycloalkenyl, heteroaryl and heterocyclic;

n is 0, 1 or 2; and

x is 1 or 2;

or a pharmaceutically acceptable salt, stereoisomer, or prodrug thereof;

provided at least one of R³ and R⁵ is a substituent comprising one ormore phosphono groups.

A preferred compound of the invention is a compound of formula Iwherein: R¹ is selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl, heterocyclic and —R^(a)—Y—R^(b)—(Z)_(x);or R¹ is a saccharide group optionally substituted with—R^(a)—Y—R^(b)—(Z)_(x), R^(f), —C(O)R^(f), or—C(O)—R^(a)—Y—R^(b)—(Z)_(x); R² is hydrogen or a saccharide groupoptionally substituted with —R^(a)—Y—R^(b)—(Z)_(x), R^(f), —C(O)R^(f),or —C(O)—R^(a)—Y—R^(b)—(Z)_(x); R³ is —OR^(c), —NR^(c)R^(c),—O—R^(a)—Y—R^(c)—(Z)_(x), —NR^(c)—R^(a)—Y—R^(b)—(Z)_(x), —NR^(c)R^(c),or —O—R^(c); or R³ is a nitrogen-linked, oxygen-linked, or sulfur-linkedsubstituent that comprises one or more phosphono groups; R⁴ is selectedfrom the group consisting of hydrogen, alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,—R^(a)—Y—R^(b)—(Z)_(x), —C(O)R^(d) and a saccharide group optionallysubstituted with —R^(a)—Y—R^(b)—(Z)_(x), R^(f), —C(O)R^(f), or—C(O)—R^(a)—Y—R^(b)—(Z)_(x); R⁵ is selected from the group consisting ofhydrogen, halo, —CH(R^(c))—NR^(c)R^(c), —CH(R^(c))—NR^(c),—CH(R^(c))—NR^(c)—R^(a)—Y—R^(b)—(Z)_(x), —CH(R^(c))—R^(x),—CH(R^(c))—NR^(c)—R^(a)—C(═O)—R^(x), and a substituent that comprisesone or more phosphono groups; R⁶ is selected from the group consistingof hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl,alkynyl, substituted alkynyl, —R^(a)—Y—R^(b)—(Z)_(x), —C(O)R^(c) and asaccharide group optionally substituted with—NR^(c)—R^(a)—Y—R^(b)—(Z)_(x), or R⁵ and R⁶ can be joined, together withthe atoms to which they are attached, form a heterocyclic ringoptionally substituted with —NR^(c)—R^(a)—Y—R^(b)—(Z)_(x); R⁷ isselected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,—R^(a)—Y—R^(b)—(Z)_(x), and —C(O)R^(d); R⁸ is selected from the groupconsisting of hydrogen, alkyl, substituted alkyl, alkenyl, substitutedalkenyl, alkynyl, substituted alkynyl, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl andheterocyclic; R⁹ is selected from the group consisting of hydrogen,alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, heteroaryl and heterocyclic; R¹⁰ isselected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic; or R⁸ and R¹⁰ arejoined to form —Ar¹—O—Ar—, where Ar¹ and Ar² are independently aryleneor heteroarylene; R¹¹ is selected from the group consisting of hydrogen,alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, heteroaryl and heterocyclic, or R¹⁰ andR¹¹ are joined, together with the carbon and nitrogen atoms to whichthey are attached, to form a heterocyclic ring; R¹² is selected from thegroup consisting of hydrogen, alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,heteroaryl, heterocyclic, —C(O)R^(d), —C(NH)R^(d), —C(O)NR^(c)R^(c),—C(O)OR^(d), —C(NH)NR^(c)R^(c) and —R^(a)—Y—R^(b)—(Z)_(x), or R¹¹ andR¹² are joined, together with the nitrogen atom to which they areattached, to form a heterocyclic ring; R¹³ is selected from the groupconsisting of hydrogen or —OR¹⁴; R¹⁴ is selected from hydrogen,—C(O)R^(d) and a saccharide group; each R^(a) is independently selectedfrom the group consisting of alkylene, substituted alkylene, alkenylene,substituted alkenylene, alkynylene and substituted alkynylene; eachR^(b) is independently selected from the group consisting of a covalentbond, alkylene, substituted alkylene, alkenylene, substitutedalkenylene, alkynylene and substituted alkynylene, provided R^(b) is nota covalent bond when Z is hydrogen; each R^(c) is independently selectedfrom the group consisting of hydrogen, alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,heteroaryl, heterocyclic and —C(O)R^(d); each R^(d) is independentlyselected from the group consisting of alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,heteroaryl and heterocyclic; R^(c) is a saccharide group; each R^(f) isindependently alkyl, substituted alkyl, alkenyl, substituted alkenyl,alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl, orheterocyclic; R^(x) is an N-linked amino saccharide or an N-linkedheterocycle; X¹, X² and X³ are independently selected from hydrogen orchloro; each Y is independently selected from the group consisting ofoxygen, sulfur, —S—S—, —NR^(c)—, —S(O)—, —SO₂—, —NR^(c)C(O)—, —OSO₂—,—OC(O)—, —NR^(c)SO₂—, —C(O)NR^(c)—, —C(O)O—, —SO₂NR^(c)—, —SO₂₀—,—P(O)(OR^(c))O—, —P(O)(OR^(c))NR^(c)—, —OP(O)(OR^(c))O—,—OP(O)(OR^(c))NR^(c)—, —OC(O)O—, —NR^(c)C(O)O—, —NR^(c)C(O)NR^(c)—,—OC(O)NR^(c)—, —C(═O)—, and —NR^(c)SO₂NR^(c)—; each Z is independentlyselected from hydrogen, aryl, cycloalkyl, cycloalkenyl, heteroaryl andheterocyclic; n is 0, 1 or 2; and x is 1 or 2; or a pharmaceuticallyacceptable salt, stereoisomer, or prodrug thereof; provided at least oneof R³ and R⁵ is a substituent comprising one or more phosphono groups.

Preferably, R¹ is a saccharide group optionally substituted with—R^(a)—Y—R^(b)—(Z)_(x), R^(f), —C(O)R^(f), or —C(O)—R^(a)—Y—R^(b)—(Z).More preferably R¹ is a saccharide group substituted on the saccharidenitrogen with —CH₂CH₂—NH—(CH₂)₉CH₃; —CH₂CH₂CH₂—NH—(CH₂)₉CH₃;—CH₂CH₂CH₂CH₂—NH—(CH₂)₇CH₃; —CH₂CH₂—NHSO₂—(CH₂)₉CH₃;—CH₂CH₂—NHSO₂—(CH₂)₁₀CH₃; —CH₂CH₂—S—(CH₂)₈CH₃; —CH₂CH₂—S—(CH₂)₉CH₃;—CH₂CH₂—S—(CH₂)₁₀CH₃; —CH₂CH₂CH₂—S—(CH₂)₈CH₃; —CH₂CH₂CH₂—S—(CH₂)₉CH₃;—CH₂CH₂CH₂—S—(CH₂)₃—CH═CH—(CH₂)₄CH₃ (trans); —CH₂CH₂CH₂CH₂—S—(CH₂)₇CH₃;—CH₂CH₂—S(O)—(CH₂)₉CH₃; —CH₂CH₂—S—(CH₂)₆Ph; —CH₂CH₂—S—(CH₂)₈Ph;—CH₂CH₂CH₂—S—(CH₂)₉Ph; —CH₂CH₂—NH—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂—NH—CH₂-4-[4-(CH₃)₂CHCH₂-]-Ph; —CH₂CH₂—NH—CH₂₄-(4-CF₃-Ph)-Ph;—CH₂CH₂—S—CH₂-4-(4-Cl-Ph)-Ph; —CH₂CH₂—S(O)—CH₂₄-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—S—CH₂-4-(4-Cl-Ph)-Ph; —CH₂CH₂CH₂—S(O)—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—S—CH₂-4-[3,4-di-Cl-PhCH₂O—)-Ph;—CH₂CH₂—NHSO₂—CH₂-4-[4-(4-Ph)-Ph]-Ph;—CH₂CH₂CH₂CH₂—NHSO₂—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—NHSO₂—CH₂-4-(Ph-C≡C—)-Ph; —CH₂CH₂CH₂—NHSO₂-4-4-Cl-Ph)-Ph; or—CH₂CH₂CH₂—NHSO₂-4-(naphth-2-yl)-Ph. Preferably R¹ is also a saccharidegroup substituted on the saccharide nitrogen with a4-(4-chlorophenyl)benzyl group or with a 4-(4-chlorobenzyloxy)benzylgroup.

In a preferred embodiment, R¹ is a saccharide group of the formula:

wherein R¹⁵ is —R^(a)—Y—R^(b)—(Z)_(x), R^(f), —C(O)R^(f), or—C(O)—R^(a)—Y—R^(b)—(Z)_(x); and R¹⁶ is hydrogen or methyl.

Preferably, R¹⁵ is —CH₂CH₂—NH—(CH₂)₉CH₃; —CH₂CH₂CH₂—NH—(CH₂)₈CH₃;—CH₂CH₂CH₂CH₂—NH— (CH₂)₇CH₃; —CH₂CH₂—NHSO₂—(CH₂)₉CH₃;—CH₂CH₂—NHSO₂—(CH₂)₁₁CH₃; —CH₂CH₂—S— (CH₂)₈CH₃; —CH₂CH₂—S— (CH₂)₉CH₃;—CH₂CH₂—S—(CH₂)₁₀CH₃; —CH₂CH₂CH₂—S— (CH₂)₈CH₃; —CH₂CH₂CH₂—S— (CH₂)₉CH₃;—CH₂CH₂CH₂—S—(CH₂)₃—CH═CH—(CH₂)₄—CH₃ (trans); —CH₂CH₂CH₂CH₂—S—(CH₂)₇CH₃; —CH₂CH₂—S(O)—(CH₂)₉CH₃; —CH₂CH₂—S—(CH₂)₆Ph;—CH₂CH₂—S—(CH₂)₈Ph; —CH₂CH₂CH₂—S— (CH₂)₈Ph;—CH₂CH₂—NH—CH₂₄-(4-Cl-Ph)-Ph; —CH₂CH₂—NH—CH₂-4-[4-(CH₃)₂CHCH₂-]-Ph;—CH₂CH₂—NH—CH₂₄-(4-CF₃-Ph)-Ph; —CH₂CH₂—S—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂—S(O)—CH₂-4-(4-Cl-Ph)-Ph; —CH₂CH₂CH₂—S—CH₂₄-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—S(O)—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—S—CH₂-4-[3,4-di-Cl-PhCH₂O—)-Ph;—CH₂CH₂—NHSO₂—CH₂-4-[4-(4-Ph)-Ph]-Ph;—CH₂CH₂CH₂—NHSO₂—CH₂₄-(4-Cl-Ph)-Ph; —CH₂CH₂CH₂—NHSO₂—CH₂-4-(Ph-C≡C—)-Ph;—CH₂CH₂CH₂—NHSO₂-4-(4-Cl-Ph)-Ph; or —CH₂CH₂CH₂—NHSO₂-4-(naphth-2-yl)-Ph.Preferably R¹⁵ can also be a 4-(4-chlorophenyl)benzyl group or a4-(4-chlorobenzyloxy)benzyl group.

Preferably, R² is hydrogen.

Preferably, R³ is —OR^(c); —NR^(c)R^(c); or a nitrogen-linked,oxygen-linked, or sulfur-linked substituent comprising one or twophosphono groups, or a pharmaceutically acceptable salt thereof. When R³is a phosphono-containing substituent, R³ is preferably anitrogen-linked substituent comprising one phosphono group, or apharmaceutically acceptable salt thereof. Preferably, R³ is a group ofthe formula —O—R^(a)— P(O)(OH)₂, —S—R^(a)—P(O)(OH)₂, or—NR^(c)—R^(a)—P(O)(OH)₂. More preferably, R³ is a group of the formula—NH—R^(a)—P(O)(OH)₂, where R^(a) is as defined herein. In this formula,R^(a) is preferably an alkylene group. Particularly preferred R³substituents include phosphonomethylamino, 3-phosphonopropylamino and2-hydroxy-2-phosphonoethylamino groups and the like.

Preferably, when R³ is not a phosphono-containing substituent, R³ is—OH; —NH—(CH₂)₃—N(CH₃)₂; N-(D-glucosamine); —NHCH(CO₂CH₃)CH₂CO₂CH₃;—NH(CH₂)₃-(morpholin-4-yl); —NH(CH₂)₃—NH(CH₂)₂CH₃;—NH(CH₂-piperidin-1-yl; —NH(CH₂)₄NHC(N)NH₂; —NH(CH₂)₂—N⁺(CH₃)₃;—NHCH(COOH)(CH₂)₃NHC(N)NH₂; —NH—[CH₂CH₂CH₂—NH-]₃—H; —N[(CH₂)₃N(CH₃)₂]₂;—NH(CH₂)₃-imidazol-1-yl; —NHCH₂-4-pyridyl; —NH(CH₂)₃CH₃; —NH(CH₂)₂OH;—NH(CH₂)₅OH; —NH(CH₂)₂OCH₃; —NHCH₂-tetrahydrofuran-2-yl; —N[(CH₂)₂OH]₂;—NH(CH₂)₂N[(CH₂)₂OH]₂; —NHCH₂COOH; —NHCH(COOH)CH₂OH; —NH(CH₂)₂COOH;N-(glucamine); —NH(CH₂)₂COOH; —NH(CH₂)₃SO₃H; —NHCH(COOH)(CH₂)₂NH₂;—NHCH(COOH)(CH₂)₃NH₂; —NHCH(COOH)CH₂CO₂(CH₂)₃—N⁺(CH₃)₃;—NHCH(COOH)CH₂CO₂(CH₂)₂C(O)—N(CH₃)₂;—NHCH(COOH)CH₂CO₂(CH₂)₃-morpholin-4-yl;—NHCH(COOH)CH₂CO₂(CH₂)₂OC(O)C(CH₃)₃; —NHCH(CH₂COOH)CO₂(CH₂)₃—N⁺(CH₃)₃;—NHCH(CH₂COOH)CO₂(CH₂)₂C(O)N(CH₃)₂; or—NHCH(CH₂COOH)CO₂(CH₂)₃-morpholin-4-yl.—NHCH(CH₂COOH)CO₂(CH₂)₂OC(O)C(CH₃)₃; —NHCH(COOH)CH₂CO₂CH₃;—NHCH(CH₂COOH)CO₂(CH₂)₂N(CH₃)₂; —NHCH(COOH)CH₂CO₂CH₂C(O)N(CH₃)₂;—NHCH(CH₂COOH)CO₂CH₂C(O)N(CH₃)₂; —NHCH(CH₂COOH)CO₂CH₃; —NH(CH₂)₃N(CH₃)₂;—NHCH₂CH₂CO₂CH₃; —NHCH[CH₂CO₂CH₂C(O)N(CH₃)₂]CO₂CH₂—C(O)—N(CH₃)₂;—NHCH₂CO₂CH₃; —N-(methyl 3-amino-3-deoxyaminopyranoside); —N-(methyl3-amino-2,3,6-trideoxyhexopyranoside);—N-(2-amino-2-deoxy-6-(dihydrogenphosphate)glucopyranose;—N-(2-amino-2-deoxygluconic acid); —NH(CH₂)₄COOH; —N—(N—CH₃-D-glucanine;—NH(CH₂)₆COOH; —O(D-glucose); —NH(CH₂)₃OC(O)CH(NH₂)CH₃;—NH(CH₂)₄—CH(C(O)₂—HOOC-pyrrolidin-1-yl)NHCH(COOH)—CH₂CH₂Ph (S,Sisomer); —NH—CH₂CH₂—NH—(CH₂)₉CH₃; —NH(CH₂)C(O)CH₂C(O)N(CH₃)₂;

Preferably, R⁴, R⁶ and R⁷ are each independently selected from hydrogenor —C(O)R^(d). More preferably, R⁴, R⁶ and R⁷ are each hydrogen.

Preferably, R⁵ is hydrogen, —CH₂—NHR^(c), —CH₂—NR^(c)R^(c),—CH₂—NH—R^(a)—Y—R^(b)—(Z)_(x), or a substituent comprising one or twophosphono groups. When R⁵ is a substituent comprising a phosphono group,R⁵ is preferably a group of the formula—CH(R²¹)—N(R^(c))—R^(a)—P(O)(OH)₂ wherein R²¹ is hydrogen or R^(d),preferably hydrogen, and R^(a), R^(c), and R^(d), are as defined herein.More preferably, when R⁵ is phosphono-containing substituent, R⁵ ispreferably a group of the formula —CH₂—NH—R^(c)—P(O)(OH)₂, where R^(a)is as defined herein. In this formula, R^(a) is preferably an alkylenegroup; more preferably, an alkylene group containing from 2 to about 6carbon atoms.

Particularly preferred R⁵ substituents includeN-(phosphonomethyl)-aminomethyl;N-(2-hydroxy-2-phosphonoethyl)aminomethyl;N-carboxymethyl-N-(2-phosphonoethyl)aminomethyl;N,N-bis(phosphonomethyl)-aminomethyl; andN-(3-phosphonopropyl)aminomethyl; and the like.

Preferably, when R⁵ is not a phosphono-containing Substituent, R⁵ ishydrogen, —CH₂—NHR^(c), —CH₂—NR^(c)R^(c) or—CH₂—NH—R^(a)—Y—R^(b)—(Z)_(x). R⁵ can also preferably be hydrogen;—CH₂—N—(N—CH₃-D-glucamine); —CH₂—NH—CH₂CH₂—NH—(CH₂)₉CH₃;—CH₂—NH—CH₂CH₂—NHC(O)—(CH₂)₃COOH; —CH₂—NH—(CH₂)₉CH₃;—CH₂—NH—CH₂CH₂—COOH; —CH₂—NH—(CH₂)₅COOH; —CH₂— (morpholin-4-yl);—CH₂—NH—CH₂CH₂—O—CH₂CH₂OH; —CH₂—NH—CH₂CH(OH)—CH₂OH; —CH₂—N[CH₂CH₂OH]₂;—CH₂—NH—(CH₂)₃—N(CH₃)₂; —CH₂—N[(CH₂)₃—N(CH₃)₂]₂;—CH₂—NH—(CH₂)₃-(imidazol-1-yl); —CH₂—NH—(CH₂)₃-(morpholin-4-yl);—CH₂—NH—(CH₂)₄—NHC(NH)NH₂; —CH₂—N-(2-amino-2-deoxygluconic acid);—CH₂—NH—CH₂CH₂—NH—(CH₂)₁₁CH₃; —CH₂—NH—CH(COOH)CH₂COOH;—CH₂—NH—CH₂CH₂—NHSO₂—(CH₂)₇CH₃; —CH₂—NH—CH₂CH₂—NHSO₂—(CH₂)₇CH₃;—CH₂—NH—CH₂CH₂—NHSO₂—(CH₂)₉CH₃; —CH₂—NH—CH₂CH₂—NHSO₂—(CH)₁₁CH₃;—CH₂—NH—CH₂CH₂—NH—(CH₂)₇CH₃; —CH₂—NH—CH₂CH₂—O—CH₂CH₂OH;—CH₂—NH—CH₂CH₂C(O)—N-(D-glucosamine);—CH₂—NH-(6-oxo-[1,3]oxazinan-3-yl); —CH₂—NH—CH₂CH₂—S—(CH₂)₇CH₃;—CH₂—NH—CH₂CH₂—S—(CH₂)₈CH₃; —CH₂—NH—CH₂CH₂—S—(CH₂)₉CH₃;—CH₂—NH—CH₂CH₂—S—CH₂)₁₁CH₃; —CH₂—NH—CH₂CH₂—S—(CH)₆Ph;—CH₂—NH—CH₂CH₂—S—(CH₂)₈Ph; —CH₂—NH—CH₂CH₂—S—(CH₂)₁₁Ph;—CH₂—NH—CH₂CH₂—S—CH₂-(4-(4-CF₃-Ph)Ph); —CH₂—NH—CH₂CH₂—NH—(CH₂)₁₁CH₃; or—CH₂—NH—(CH₂)₅—COOH.

Preferably, R⁸ is —CH₂C(O)NH₂, —CH₂COOH, benzyl, 4-hydroxyphenyl or3-chloro-4-hydroxyphenyl.

Preferably, R⁹ is hydrogen or alkyl.

Preferably, R¹⁰ is alkyl or substituted alkyl More preferably, R¹⁰ isthe side-chain of a naturally occurring amino acid, such as isobutyl.

Preferably, R¹¹ is hydrogen or alkyl.

Preferably, R¹² is hydrogen, alkyl, substituted alkyl or —C(O)R^(d). R¹²can also preferably be hydrogen; —CH₂COOH; —CH₂—[CH(OH)]₅CH₂OH;—CH₂CH(OH)CH₂OH; —CH₂CH₂NH₂; —CH₂C(O)OCH₂CH₃; —CH₂-(2-pyridyl);—CH₂—[CH(OH)]₄COOH; —CH₂-(3-carboxyphenyl); (R)—C(O)CH(NH₂)(CH₂)₄NH₂;—C(O)Ph; —C(O)CH₂NHC(O)CH₃; E-CH₂CH₂—S—(CH₂)₃CH═CH(CH)₄CH₃; or —C(O)CH₃.

Preferably, X¹ and X² are each chloro.

Preferably, X³ is hydrogen.

Preferably, each Y is independently selected from the group consistingof oxygen, sulfur, —S—S—, —NR^(c)—, —S(O)—, —SO₂—, —NR^(c)(O)—, —OSO₂—,—OC(O)—, —NR^(c)SO₂—, —C(O)NR^(c)—, —C(O)O—, —SO₂NR^(c)—, —SO₂O—,—P(O)(OR^(c))O—, —P(O)(OR^(c))NR^(c)—, —OP(O)(OR^(c))O—,—OP(O)(OR^(c))NR^(c)—, —OC(O)O—, —NR^(c)C(O)O—, —NR^(c)(O)NR^(c)—,—OC(O)NR^(c)—, and —NR^(c)SO₂NR^(c)—.

Preferably, n is 0 or 1, and more preferably, n is 1.

Another preferred compound of the invention is a glycopeptide of formulaII:

wherein:

R⁹ is hydrogen;

R²⁰ is —R^(a)—Y—R^(b)—(Z)_(x), R^(f), —C(O)R^(f), or—C(O)—R^(a)—Y—R^(b)—(Z)_(x); and

R^(a), Y, R^(b), Z, x, R^(f), R³, and R⁵ have any of the values orpreferred values described herein;

or a pharmaceutically acceptable salt, stereoisomer, or prodrug thereof;

provided at least one of R³ and R⁵ is a substituent comprising one ormore phosphono groups.

Preferably, R²⁰ is —CH₂CH₂—NH—(CH₂)₉CH₃; —CH₂CH₂CH₂—NH—(CH)₉CH₃;—CH₂CH₂CH₂CH₂—NH—(CH)₇CH₃; —CH₂CH₂—NHSO₂—(CH₂)₉CH₃;—CH₂CH₂—NHSO₂—(CH₂)₁₁CH₃; —CH₂CH₂—S—(CH₂)₈CH₃; —CH₂CH₂—S— (CH₂)₉CH₃;—CH₂CH₂—S—(CH₂)₁₀CH₃; —CH₂CH₂CH₂—S—(CH₂)₈CH₃; —CH₂CH₂CH₂—S— (CH₂)₉CH₃;—CH₂CH₂CH₂—S— (CH₂)₃—CH═CH—(CH₂)₄—CH₃ (trans); —CH₂CH₂CH₂CH₂—S—(CH₂)₇CH₃; —CH₂CH₂—S(O)—(CH₂)₉CH₃; —CH₂CH₂—S—(CH₂)₆Ph;—CH₂CH₂—S—(CH₂)₈Ph; —CH₂CH₂CH₂—S—(CH₂)₈Ph;—CH₂CH₂—NH—CH₂-4-(4-Cl-Ph)-Ph; —CH₂CH₂—NH—CH₂-4-[4-(CH₃)₂CHCH₂—]-Ph;—CH₂CH₂—NH—CH₂-4-(4-CF₃-Ph)-Ph; —CH₂CH₂—S—CH₂₄-(4-Cl-Ph)-Ph;—CH₂CH₂—S(O)—CH₂₄-(4-Cl-Ph)-Ph; —CH₂CH₂CH₂—S—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—S(O)—CH₂₄-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—S—CH₂-4-[3,4-di-Cl-PhCH₂O—)-Ph;—CH₂CH₂—NHSO₂—CH₂-4-[4-(4-Ph)-Ph]-Ph;—CH₂CH₂CH₂—NHSO₂—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—NHSO₂—CH₂-4-(Ph-C≡C—)-Ph; —CH₂CH₂CH₂—NHSO₂-4-(4-Cl-Ph)-Ph; or—CH₂CH₂CH₂—NHSO₂-4-(naphth-2-yl)-Ph. Preferably R²⁰ is also a4-(4-chlorophenyl)benzyl group or a 4-(4-chlorobenzyloxy)benzyl group.

In another preferred embodiment, the invention provides a compound offormula II, wherein R¹⁹ is hydrogen; R²⁰ is —CH₂CH₂NH—CH₂)₉CH₃; R³ is—OH; and R⁵ is a substituent comprising a phosphono group; or apharmaceutically acceptable salt thereof.

In yet another preferred embodiment, the invention provides a compoundof formula II, wherein R¹⁹ is hydrogen; R²⁰ is —R^(a)—Y—R^(b)—(Z)_(x),R^(f), —C(O)R^(f), or —C(O)—R^(a)—Y—R^(b)—(Z)_(x); R³ is —OH; and R⁵ is—CH₂—NH—CH₂—P(O)(OH)₂; or a pharmaceutically acceptable salt thereof.

The invention also provides a pharmaceutical composition comprising apharmaceutically acceptable carrier and a therapeutically effectiveamount of a compound of the invention. In one preferred embodiment, thepharmaceutically acceptable carrier comprises an aqueous cyclodextrinsolution. Preferably, the cyclodextrin is hydroxypropyl-β-cyclodextrinor sulfobutyl ether β-cyclodextrin More preferably, the cyclodextrin ishydroxypropyl-β-cyclodextrin.

The compounds of the invention are highly effective antibacterialagents. Accordingly, the invention also provides a method of treating amammal having a bacterial disease, comprising administering to themammal a therapeutically effective amount of a compound of theinvention. The invention also provides a method of treating a mammalhaving a bacterial disease, comprising administering to the mammal atherapeutically effective amount of a pharmaceutical composition of theinvention.

The invention also provides processes and intermediates useful forpreparing compounds of the invention, which processes and intermediatesare described further herein.

The invention also provides a compound of the invention as describedherein for use in medical therapy, as well as the use of a compound ofthe invention in the manufacture of a formulation or medicament fortreating a bacterial disease in a mammal.

The invention also provides a pharmaceutical composition which comprisesas an active ingredient a compound of the invention for the treatment ofa bacterial disease.

The invention also provides a method for preparing a glycopeptide of theinvention which is substituted at the C-terminus with a substituent thatcomprises one or more phosphono groups, comprising coupling acorresponding starting glycopeptide wherein the C-terminus is a carboxygroup with a suitable phosphono containing compound.

The invention also provides a method for preparing a glycopeptide of theinvention which is substituted at the R-terminus with a substituent thatcomprises one or more phosphono groups, comprising coupling acorresponding starting glycopeptide wherein the R-terminus isunsubstituted with a suitable phosphono containing compound. When thestarting glycopeptide is substituted at the vancosamine amino terminus,such a method can further optionally comprise preparing the startingglycopeptide by reductively alkylating a corresponding glycopeptidewherein the vancosamine amino terminus is the corresponding amine.

The invention also provides a method for preparing a glycopeptide of theinvention that is substituted at the C-terminus, comprising derivatizinga corresponding starting glycopeptide wherein the C-terminus is acarboxy group.

The invention also provides a method for preparing a glycopeptide of theinvention which is substituted at the R-terminus, comprisingderivatizing a corresponding starting glycopeptide wherein theR-terminus is unsubstituted (i.e. a hydrogen)

This invention also provides a method for preparing a compound offormula II, wherein R³ is —OH, R⁵ is —CH₂—NH—R^(c)—P(O)(OH)₂, R¹⁹ ishydrogen and R²⁰ is —R^(a)—Y—R^(b)—(Z)_(x), or —R^(f), and R^(a), R^(b),R^(f), Y, Z and x are as defined herein, or salt thereof; the methodcomprising:

(a) reductively alkylating a compound of formula II, wherein R³ is —OHand R⁵, R¹⁹ and R²⁰ are hydrogen, or a salt thereof, with an aldehyde ofthe formula HC(O)—R^(a′)—Y—R^(b)—(Z)_(x) or HC(O)R^(f′) wherein R^(a′)and R^(f′) represent R^(a) and R^(f), respectively, minus one —CH₂—group, to form a compound of formula II wherein R³ is —OH, R⁵ and R¹⁹are hydrogen and R²⁰ is —R^(a)—Y—R^(b)—(Z)_(x) or —R^(f), or saltthereof; and

(b) reacting the product from step (a) with formaldehyde andH₂N—R^(a)—P(O)(OH)₂ to form a compound of formula II wherein R³ is —OH,R⁵ is —CH₂NH—R^(a)—P(O)(OH)₂, R¹⁹ is hydrogen and R²⁰ is—R^(a)—Y—R^(b)—(Z)_(x) or —R^(f), or salt thereof.

Preferred compounds of the invention are the compounds of formula IIshown in Table I below wherein R¹⁹ is hydrogen.

TABLE I Preferred Compounds of formula II Compound R³ R⁵ R²⁰ 1phosphonomethylamino H CH₃(CH₂)₉NHCH₂CH₂— 2 phosphonomethylamino HCH₃(CH₂)₉OCH₂CH₂— 3 phosphonomethylamino H CH₃(CH₂)₉SCH₂CH₂— 4phosphonomethylamino H CH₃(CH₂)₁₂— 5 phosphonomethylamino H4-(4-chlorophenyl)-benzyl 6 phosphonomethylamino H2-(4-(4-chlorophenyl)- benzylamino)ethyl 7 phosphonomethylamino H4-(4′-chlorobiphenyl)-butyl 8 phosphonomethylamino H5-(4′-chlorobiphenyl)-pentyl 9 3-phosphonopropylamino HCH₃(CH₂)₉SCH₂CH₂— 10 2-hydroxy-2- H 4-(4-chlorophenyl)-phosphonoethylamino benzyl 11 OH (phosphonomethyl)- CH₃(CH₂)₉NHCH₂CH₂—aminomethyl 12 OH (phosphonomethyl)- CH₃(CH₂)₉SCH₂CH₂— aminomethyl 13 OH(phosphonomethyl)- CH₃(CH₂)₉OCH₂CH₂— aminomethyl 14 OH(phosphonomethyl)- CH₃(CH₂)₁₂— aminomethyl 15 OH (phosphonomethyl)-4-(4- aminomethyl chlorophenyl)benzyl 16 OH (phosphonomethyl)-2-(4-(4-chlorophenyl)- aminomethyl benzylamino)ethyl 17 OH(phosphonomethyl)- 4-(4′- aminomethyl chlorobiphenyl)butyl 18 OH(phosphonomethyl)- 5-(4′- aminomethyl chlorobiphenyl)pentyl 19 OH(phosphonomethyl)- 3-[4-(4- aminomethyl chlorobenzyloxy)-benzylthio]propyl 20 OH N-(2-hydroxy-2-phos- CH₃(CH₂)₉SCH₂CH₂—phonoethyl)aminomethyl 21 OH N-(carboxymethyl)-N-2- CH₃(CH₂)₉SCH₂CH₂—phosphonomethyl)- aminomethyl 22 OH N,N-bis(phosphono-CH₃(CH₂)₉NHCH₂CH₂— methyl)aminomethyl 23 OH 3-phosphonopropyl-CH₃(CH₂)₉SCH₂CH₂— aminomethyl 24 OH 3-phosphonopropyl-4-(4-chlorophenyl)benzyl aminomethyl 25 phosphonomethylamino—CH₂—N—(N—CH₃-D- CH₃(CH₂)₉NHCH₂CH₂— glucamine 26 OH (phosphonomethyl)-—(CH₂)₃NH—SO₂-4-(4- aminomethyl chlorophenyl)phenyl

Another preferred group of compounds of the invention are phosphonoderivatives of the glycopeptide antibiotic A82846B (also known aschloroorienticin A oy LY264826). See for example R. Nagarajan et al., J.Org. Chem., 1988, 54, 983-986; and N. Tsuji et al., J. Antibiot., 1988,41, 819-822. The structure of this glycopeptide is similar tovancomycin, except A82846B contains an additional amino sugar (i.e.4-epi-vancosamine attached at the R² position in formula I.) and furthercontains 4-epi-vancosamine in place of vancosamine in the disaccharidemoiety attached at the R¹ position in formula I. For example, apreferred group of compounds are N-alkylated derivatives of A82846B thatare substituted at the C-terminus or the R-terminus with a substituentthat comprises one or more (e.g. 1, 2, 3, 4, or 5) phosphono (—PO₃H₂)groups; or a pharmaceutically acceptable salt thereof. A preferred groupof compounds of the invention that are derivatives of A82846B aresubstituted at either the C-terminus or the R-terminus with asubstituent that comprises one or more (e.g. 1, 2, 3, 4, or 5) phosphono(—PO₃H₂) groups. Another preferred group of compounds of the inventionthat are derivatives of A82846B are substituted at the C-terminus andthe R-terminus with substituents that each comprises one or more (e.g.1, 2, 3, 4, or 5) phosphono (—PO₃H₂) groups. Another preferred group ofcompounds of the invention are phosphono derivatives of A82846B having a4-(4-chlorophenyl)benzyl group or a 4-(4-chlorobenzyloxy)benzyl groupattached at the amino group of the 4-epi-vancosamine of the disaccharidemoiety. The compounds of the invention that are phosphono derivatives ofA82846B can readily be prepared using the procedures described herein.

The phosphono compounds of the invention have been found to unexpectedlyexhibit reduced tissue accumulation and/or nephrotoxicity whenadministered to a mammal. While not wishing to be bound by theory, it isbelieved that the phosphono moiety serves to increase the overallnegative charge of the glycopeptide under physiological conditionsthereby facilitating excretion from the mammal after administration. Theunexpected increase in excretion of the phosphono compounds of theinvention may be responsible for the reduced tissue accumulation and/orreduced nephrotoxicity observed for these compounds relative to thecorresponding compounds that lack the phosphono functionality.

DETAILED DESCRIPTION OF THE INVENTION

This invention relates to novel compounds of the invention, which arederivatives of glycopeptide antibiotics comprising one or moresubstituents that comprise one or more phosphono groups, as well as tocompositions comprising such compounds and to therapeutic methodscomprising the administration of such compounds. When describing thecompounds, compositions and methods of the invention, the followingterms have the following meanings, unless otherwise indicated.

DEFINITIONS

The term “alkyl” refers to a monoradical branched or unbranchedsaturated hydrocarbon chain preferably having from 1 to 40 carbon atoms,more preferably 1 to 10 carbon atoms, and even more preferably 1 to 6carbon atoms. This term is exemplified by groups such as methyl, ethyl,n-propyl, iso-propyl, n-butyl, iso-butyl, n-hexyl, n-decyl, tetradecyl,and the like.

The term “substituted alkyl” refers to an alkyl group as defined above,having from 1 to 8 substituents, preferably 1 to 5 substituents, andmore preferably 1 to 3 substituents, selected from the group consistingof alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino,substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano,halogen, hydroxyl, keto, thioketo, carboxy, carboxyalkyl, thioaryloxy,thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substitutedthioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclic,heterocyclooxy, hydroxyamino, alkoxyamino, nitro, —SO-alkyl,—SO-substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl, —SO₃H, guanido, and —SO₂-heteroaryl.

The term “alkylene” refers to a diradical of a branched or unbranchedsaturated hydrocarbon chain, preferably having from 1 to 40 carbonatoms, preferably 1-10 carbon atoms, more preferably 1-6 carbon atoms.This term is exemplified by groups such as methylene (—CH₂—), ethylene(—CH₂CH₂—), the propylene isomers (e.g., —CH₂CH₂CH₂— and —CH(CH₁)CH₂—)and the like.

The term “substituted alkylene” refers to an alkylene group, as definedabove, having from 1 to 5 substituents, and preferably 1 to 3substituents, selected from the group consisting of alkoxy, substitutedalkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryland —SO₂-heteroaryl. Additionally, such substituted alkylene groupsinclude those where 2 substituents on the alkylene group are fused toform one or more cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, heterocyclic or heteroaryl groups fusedto the alkylene group. Preferably such fused groups contain from 1 to 3fused ring structures. Additionally, the term substituted alkyleneincludes alkylene groups in which from 1 to 5 of the alkylene carbonatoms are replaced with oxygen, sulfur or —NR— where R is hydrogen oralkyl. Examples of substituted alkylenes are chloromethylene (—CH(Cl)—),aminoethylene (—CH(NH₂)CH₂—), 2-carboxypropylene isomers(—CH₂CH(CO₂H)CH₂—), ethoxyethyl (—CH₂CH₂O—CH₂CH₂—) and the like.

The term “alkaryl” refers to the groups -alkylene-aryl and -substitutedalkylene-aryl where alkylene, substituted alkylene and aryl are definedherein. Such alkaryl groups are exemplified by benzyl, phenethyl and thelike.

The term “alkoxy” refers to the groups alkyl-O—, alkenyl-O—,cycloalkyl-O—, cycloalkenyl-O—, and alkynyl-O—, where alkyl, alkenyl,cycloalkyl, cycloalkenyl, and alkynyl are as defined herein. Preferredalkoxy groups are alkyl-O— and include, by way of example, methoxy,ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy, sec-butoxy,n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like.

The term “substituted alkoxy” refers to the groups substituted alkyl-O—,substituted alkenyl-O—, substituted cycloalkyl-O—, substitutedcycloalkenyl-O—, and substituted alkynyl-O— where substituted alkyl,substituted alkenyl, substituted cycloalkyl, substituted cycloalkenyland substituted alkynyl are as defined herein.

The term “alkylalkoxy” refers to the groups -alkylene-O-alkyl,alkylene-O-substituted alkyl, substituted alkylene-O-alkyl andsubstituted alkylene-O-substituted alkyl wherein alkyl, substitutedalkyl, alkylene and substituted alkylene are as defined herein.Preferred alkylalkoxy groups are alkylene-O-alkyl and include, by way ofexample, methylenemethoxy (—CH₂OCH₃), ethylenemethoxy (—CH₂CH₂OCH₃),n-propylene-iso-propoxy (—CH₂CH₂CH₂OCH(CH₃)₂), methylene-t-butoxy(—CH₂—O—C(CH₃)₃) and the like.

The term “alkylthioalkoxy” refers to the group -alkylene-5-alkyl,alkylene-5-substituted alkyl, substituted alkylene-5-alkyl andsubstituted alkylene-S-substituted alkyl wherein alkyl, substitutedalkyl, alkylene and substituted alkylene are as defined herein.Preferred alkylthioalkoxy groups are alkylene-5-alkyl and include, byway of example, methylenethiomethoxy (—CH₂SCH₃), ethylenethiomethoxy(—CH₂CH₂SCH₃), n-propylene-iso-thiopropoxy (—CH₂CH₂CH₂SCH(CH₃)₂),methylene-t-thiobutoxy

(—CH₂SC(CH₃)₃) and the like.

The term “alkenyl” refers to a monoradical of a branched or unbranchedunsaturated hydrocarbon group preferably having from 2 to 40 carbonatoms, more preferably 2 to 10 carbon atoms and even more preferably 2to 6 carbon atoms and having at least 1 and preferably from 1-6 sites ofvinyl unsaturation. Preferred alkenyl groups include ethenyl (—CH═CH₂),n-propenyl (—CH₂CH═CH₂), iso-propenyl (—C(CH₃)═CH₂), and the like.

The term “substituted alkenyl” refers to an alkenyl group as definedabove having from 1 to 5 substituents, and preferably 1 to 3substituents, selected from the group consisting of alkoxy, substitutedalkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,keto, thioketo, carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryland —SO₂-heteroaryl.

The term “alkenylene” refers to a diradical of a branched or unbranchedunsaturated hydrocarbon group preferably having from 2 to 40 carbonatoms, more preferably 2 to 10 carbon atoms and even more preferably 2to 6 carbon atoms and having at least 1 and preferably from 1-6 sites ofvinyl unsaturation. This term is exemplified by groups such asethenylene (—CH═CH—), the propenylene isomers (e.g., —CH₂CH═CH— and—C(CH₃)═CH—) and the like.

The term “substituted alkenylene” refers to an alkenylene group asdefined above having from 1 to 5 substituents, and preferably from 1 to3 substituents, selected from the group consisting of alkoxy,substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substitutedamino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen,hydroxyl, carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryland —SO₂-heteroaryl. Additionally, such substituted alkenylene groupsinclude those where 2 substituents on the alkenylene group are fused toform one or more cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, heterocyclic or heteroaryl groups fusedto the alkenylene group.

The term “alkynyl” refers to a monoradical of an unsaturated hydrocarbonpreferably having from 2 to 40 carbon atoms, more preferably 2 to 20carbon atoms and even more preferably 2 to 6 carbon atoms and having atleast 1 and preferably from 1-6 sites of acetylene (triple bond)unsaturation. Preferred alkynyl groups include ethynyl (—C≡CH),propargyl (—CH₂C≡CH) and the like.

The term “substituted alkynyl” refers to an alkynyl group as definedabove having from 1 to 5 substituents, and preferably 1 to 3substituents, selected from the group consisting of alkoxy, substitutedalkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryland —SO₂-heteroaryl.

The term “alkynylene” refers to a diradical of an unsaturatedhydrocarbon preferably having from 2 to 40 carbon atoms, more preferably2 to 10 carbon atoms and even more preferably 2 to 6 carbon atoms andhaving at least 1 and preferably from 1-6 sites of acetylene (triplebond) unsaturation. Preferred alkynylene groups include ethynylene(—C≡C—), propargylene (—CH₂C≡C—) and the like.

The term “substituted alkynylene” refers to an alkynylene group asdefined above having from 1 to 5 substituents, and preferably 1 to 3substituents, selected from the group consisting of alkoxy, substitutedalkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,keto, thioketo, carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryland —SO₂-heteroaryl.

The term “acyl” refers to the groups HC(O)—, alkyl-C(O)—, substitutedalkyl-C(O)—, cycloalkyl-C(O)—, substituted cycloalkyl-C(O)—,cycloalkenyl-C(O), substituted cycloalkenyl-C(O)—, aryl-C(O)—,heteroaryl-C(O)— and heterocyclic-C(O)— where alkyl, substituted alkyl,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic are as defined herein.

The term “acylamino” or “aminocarbonyl” refers to the group —C(O)NRRwhere each R is independently hydrogen, alkyl, substituted alkyl, aryl,heteroaryl, heterocyclic or where both R groups are joined to form aheterocyclic group (e.g., morpholino) wherein alkyl, substituted alkyl,aryl, heteroaryl and heterocyclic are as defined herein.

The term “aminoacyl” refers to the group —NRC(O)R where each R isindependently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, orheterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl andheterocyclic are as defined herein.

The term “aminoacyloxy” or “alkoxycarbonylamino” refers to the group—NRC(O)OR where each R is independently hydrogen, alkyl, substitutedalkyl, aryl, heteroaryl, or heterocyclic wherein alkyl, substitutedalkyl, aryl, heteroaryl and heterocyclic are as defined herein.

The term “acyloxy” refers to the groups alkyl-C(O)O—, substitutedalkyl-C(O)O—, cycloalkyl-C(O)O—, substituted cycloalkyl-C(O)O—,aryl-C(O)O—, heteroaryl-C(O)O—, and heterocyclic-C(O)O— wherein alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, heteroaryl,and heterocyclic are as defined herein.

The term “aryl” refers to an unsaturated aromatic carbocyclic group offrom 6 to 20 carbon atoms having a single ring (e.g., phenyl) ormultiple condensed (fused) rings, wherein at least one ring is aromatic(e.g., naphthyl, dihydrophenanthrenyl, fluorenyl, or anthryl). Preferredaryls include phenyl, naphthyl and the like.

Unless otherwise constrained by the definition for the aryl substituent,such aryl groups can optionally be substituted with from 1 to 5substituents, preferably 1 to 3 substituents, selected from the groupconsisting of acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl,alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substitutedalkoxy, substituted alkenyl, substituted alkynyl, substitutedcycloalkyl, substituted cycloalkenyl, amino, substituted amino,aminoacyl, acylamino, alkaryl, aryl, aryloxy, azido, carboxy,carboxyalkyl, cyano, halo, nitro, heteroaryl, heteroaryloxy,heterocyclic, heterocyclooxy, aminoacyloxy, oxyacylamino, sulfonamide,thioalkoxy, substituted thioalkoxy, thioaryloxy, thioheteroaryloxy,—SO-alkyl, —SO-substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl, —SO₂-heteroaryl and trihalomethyl.Preferred aryl substituents include alkyl, alkoxy, halo, cyano, nitro,trihalomethyl, and thioalkoxy.

The term “aryloxy” refers to the group aryl-O— wherein the aryl group isas defined above including optionally substituted aryl groups as alsodefined above.

The term “arylene” refers to the diradical derived from aryl (includingsubstituted aryl) as defined above and is exemplified by 1,2-phenylene,1,3-phenylene, 1,4-phenylene, 1,2-naphthylene and the like.

The term “amino” refers to the group —NH₂.

The term “substituted amino” refers to the group —NRR where each R isindependently selected from the group consisting of hydrogen, alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl,substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkynyl,substituted alkynyl, aryl, heteroaryl and heterocyclic provided thatboth R's are not hydrogen.

“Amino acid” refers to any of the naturally occurring amino acids (e.g.Ala, Arg, Asn, Asp, Cys, Glu, Gln, Gly, His, Hyl, Hyp, Ile, Leu, Lys,Met, Phe, Pro, Ser, Thr, Trp, Tyr, and Val) in D, L, or DL form. Theside chains of naturally occurring amino acids are well known in the artand include, for example, hydrogen (e.g., as in glycine), alkyl (e.g.,as in alanine, valine, leucine, isoleucine, proline), substituted alkyl(e.g., as in threonine, serine, methionine, cysteine, aspartic acid,asparagine, glutamic acid, glutamine, arginine, and lysine), alkaryl(e.g., as in phenylalanine and tryptophan), substituted arylalkyl (e.g.,as in tyrosine), and heteroarylalkyl (e.g., as in histidine).

The term “carboxy” refers to —COOH.

The term “C-terminus” as it relates to a glycopeptide is well understoodin the art. For example, for a glycopeptide of formula I, the C-terminusis the position substituted by the group R³.

The term “dicarboxy-substituted alkyl” refers to an alkyl groupsubstituted with two carboxy groups. This term includes, by way ofexample, —CH₂(COOH)CH₂COOH and —CH₂(COOH)CH₂CH₂COOH.

The term “carboxyalkyl” or “alkoxycarbonyl” refers to the groups“—C(O)O-alkyl”, “—C(O)O-substituted alkyl”, “—C(O)O-cycloalkyl”,“—C(O)O-substituted cycloalkyl”, “—C(O)O-alkenyl”, “—C(O)O-substitutedalkenyl”, “—C(O)O-alkynyl” and “—C(O)O-substituted alkynyl” where alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl,substituted alkenyl, alkynyl and substituted alkynyl are as definedherein.

The term “cycloalkyl” refers to cyclic alkyl groups of from 3 to 20carbon atoms having a single cyclic ring or multiple condensed rings.Such cycloalkyl groups include, by way of example, single ringstructures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, andthe like, or multiple ring structures such as adamantanyl, and the like.

The term “substituted cycloalkyl” refers to cycloalkyl groups havingfrom 1 to 5 substituents, and preferably 1 to 3 substituents, selectedfrom the group consisting of alkoxy, substituted alkoxy, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl,acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy,oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxy,carboxyalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol,thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino,nitro, —SO-alkyl, —SO-substituted alkyl, —SO-aryl, —SO-heteroaryl,—SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl and —SO₂-heteroaryl.

The term “cycloalkenyl” refers to cyclic alkenyl groups of from 4 to 20carbon atoms having a single cyclic ring and at least one point ofinternal unsaturation. Examples of suitable cycloalkenyl groups include,for instance, cyclobut-2-enyl, cyclopent-3-enyl, cyclooct-3-enyl and thelike.

The term “substituted cycloalkenyl” refers to cycloalkenyl groups havingfrom 1 to 5 substituents, and preferably 1 to 3 substituents, selectedfrom the group consisting of alkoxy, substituted alkoxy, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl,acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy,oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxy,carboxyalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol,thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino,nitro, —SO-alkyl, —SO-substituted alkyl, —SO-aryl, —SO-heteroaryl,—SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl and —SO₂-heteroaryl.

The term “halo” or “halogen” refers to fluoro, chloro, bromo and iodo.

“Haloalkyl” refers to alkyl as defined herein substituted by 1-4 halogroups as defined herein, which may be the same or different.Representative haloalkyl groups include, by way of example,trifluoromethyl, 3-fluorododecyl, 12,12,12-trifluorododecyl,2-bromooctyl, 3-bromo-6-chloroheptyl, and the like.

The term “heteroaryl” refers to an aromatic group of from 1 to 15 carbonatoms and 1 to 4 heteroatoms selected from oxygen, nitrogen and sulfurwithin at least one ring (if there is more than one ring).

Unless otherwise constrained by the definition for the heteroarylsubstituent, such heteroaryl groups can be optionally substituted with 1to 5 substituents, preferably 1 to 3 substituents, selected from thegroup consisting of acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy,alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl,substituted alkoxy, substituted alkenyl, substituted alkynyl,substituted cycloalkyl, substituted cycloalkenyl, amino, substitutedamino, aminoacyl, acylamino, alkaryl, aryl, aryloxy, azido, carboxy,carboxyalkyl, cyano, halo, nitro, heteroaryl, heteroaryloxy,heterocyclic, heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy,substituted thioalkoxy, thioaryloxy, thioheteroaryloxy, —SO-alkyl,—SO-substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-ary, —SO₂-heteroaryl and trihalomethyl.Preferred aryl substituents include alkyl, alkoxy, halo, cyano, nitro,trihalomethyl, and thioalkoxy. Such heteroaryl groups can have a singlering (e.g., pyridyl or furyl) or multiple condensed rings (e.g.,indolizinyl or benzothienyl). Preferred heteroaryls include pyridyl,pyrrolyl and furyl.

“Heteroarylalkyl” refers to (heteroaryl)alkyl- where heteroaryl andalkyl are as defined herein. Representative examples include2-pyridylmethyl and the like.

The term “heteroaryloxy” refers to the group heteroaryl-O—.

The term “heteroarylene” refers to the diradical group derived fromheteroaryl (including substituted heteroaryl), as defined above, and isexemplified by the groups 2,6-pyridylene, 2,4-pyridiylene,1,2-quinolinylene, 1,8-quinolinylene, 1,4-benzofuranylene,2,5-pyridinylene, 2,5-indolenyl and the like.

The term “heterocycle” or “heterocyclic” refers to a monoradicalsaturated or unsaturated group having a single ring or multiplecondensed rings, from 1 to 40 carbon atoms and from 1 to 10 heteroatoms, preferably 1 to 4 heteroatoms, selected from nitrogen, sulfur,phosphorus, and/or oxygen within the ring.

Unless otherwise constrained by the definition for the heterocyclicsubstituent, such heterocyclic groups can be optionally substituted with1 to 5, and preferably 1 to 3 substituents, selected from the groupconsisting of alkoxy, substituted alkoxy, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino,acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy,oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxy,carboxyalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol,thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino,nitro, —SO-alkyl, —SO-substituted alkyl, —SO-aryl, —SO-heteroaryl,—SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl, oxo (═O), and—SO₂-heteroaryl. Such heterocyclic groups can have a single ring ormultiple condensed rings. Preferred heterocyclics include morpholino,piperidinyl, and the like.

Examples of nitrogen heterocycles and heteroaryls include, but are notlimited to, pyrrole, imidazole, pyrazole, pyridine, pyrazine,pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine,quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine,quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline,phenanthridine, acridine, phenanthroline, isothiazole, phenazine,isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline,piperidine, piperazine, indoline, morpholino, piperidinyl,tetrahydrofuranyl, and the like as well as N-alkoxy-nitrogen containingheterocycles.

Another class of heterocyclics is known as “crown compounds” whichrefers to a specific class of heterocyclic compounds having one or morerepeating units of the formula [—(CH₂—)_(a)A-] where a is equal to orgreater than 2, and A at each separate occurrence can be O, N, S or P.Examples of crown compounds include, by way of example only,[—(CH₂)₃—NH-]₃, [—((CH₂)₂—O)₄—((CH₂)₂—NH)₂] and the like. Typically suchcrown compounds can have from 4 to 10 heteroatoms and 8 to 40 carbonatoms.

The term “heterocyclooxy” refers to the group heterocyclic-O—.

The term “thioheterocyclooxy” refers to the group heterocyclic-S—.

The term “N-terminus” as it relates to a glycopeptide is well understoodin the art. For example, for a glycopeptide of formula II, theN-terminus is the position substituted by the group R¹⁹ and R²⁰.

The term “oxyacylamino” or “aminocarbonyloxy” refers to the group—OC(O)NRR where each R is independently hydrogen, alkyl, substitutedalkyl, aryl, heteroaryl, or heterocyclic wherein alkyl, substitutedalkyl, aryl, heteroaryl and heterocyclic are as defined herein.

The term “phosphono” refers to —PO₃H₂.

The term “phosphonomethylamino” refers to —NH—CH₂—P(O)(OH)₂.

The term “phosphonomethylaminomethyl” refers to —CH₂—NH—CH₂—P(O)(OH)₂.

The term “prodrug” is well understood in the art and includes compoundsthat are converted to pharmaceutically active compounds of the inventionin a mammalian system. For example, see Remington's PharmaceuticalSciences, 1980, vol. 16, Mack Publishing Company, Easton, Pa., 61 and424.

The term “R-terminus” as it relates to a glycopeptide is well understoodin the art. For example, for a glycopeptide of formula I, the R-terminusis the position substituted by the group R⁵.

The term “saccharide group” refers to an oxidized, reduced orsubstituted saccharide monoradical covalently attached to theglycopeptide or other compound via any atom of the saccharide moiety,preferably via the aglycone carbon atom. The term includesamino-containing saccharide groups. Representative saccharide include,by way of illustration, hexoses such as D-glucose, D-mannose, D-xylose,D-galactose, vancosamine, 3-desmethyl-vancosamine, 3-epi-vancosamine,4-epi-vancosanine, acosamine, actinosamine, daunosamine,3-epi-daunosamine, ristosamine, D-glucamine, N-methyl-D-glucamine,D-glucuronic acid, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine,sialyic acid, iduronic acid, L-fucose, and the like; pentoses such asD-ribose or D-arabinose; ketoses such as D-ribulose or D-fructose;disaccharides such as 2-O-(α-L-vancosaminyl)-β-D-glucopyranose,2-O-(3-desmethyl-α-L-vancosaminyl)-β-D-glucopyranose, sucrose, lactose,or maltose; derivatives such as acetals, amines, acylated, sulfated andphosphorylated sugars; oligosaccharides having from 2 to 10 saccharideunits. For the purposes of this definition, these saccharide arereferenced using conventional three letter nomenclature and thesaccharide can be either in their open or preferably in their pyranoseform.

The term “amino-containing saccharide group” refers to a saccharidegroup having an amino substituent. Representative amino-containingsaccharide include L-vancosamine, 3-desmethyl-vancosamine,3-epi-vancosamine, 4-epi-vancosamine, acosamine, actinosamine,daunosamine, 3-epi-daunosamine, ristosamine, N-methyl-D-glucamine andthe like.

The term “spiro-attached cycloalkyl group” refers to a cycloalkyl groupattached to another ring via one carbon atom common to both rings.

The term “stereoisomer” as it relates to a given compound is wellunderstood in the art, and refers another compound having the samemolecular formula, wherein the atoms making up the other compound differin the way they are oriented in space, but wherein the atoms in theother compound are like the atoms in the given compound with respect towhich atoms are joined to which other atoms (e.g. an enantiomer, adiastereomer, or a geometric isomer). See for example, Morrison andBoyde Organic Chemistry, 1983, 4th ed., Allyn and Bacon, Inc., Boston,Mass., page 123

The term “sulfonamide” refers to a group of the formula —SO₂NRR, whereeach R is independently hydrogen, alkyl, substituted alkyl, aryl,heteroaryl, or heterocyclic wherein alkyl, substituted alkyl, aryl,heteroaryl and heterocyclic are as defined herein.

The term “thiol” refers to the group —SH.

The term “thioalkoxy” refers to the group —S-alkyl.

The term “substituted thioalkoxy” refers to the group —S-substitutedalkyl.

The term “thioaryloxy” refers to the group aryl-S— wherein the arylgroup is as defined above including optionally substituted aryl groupsalso defined above.

The term “thioheteroaryloxy” refers to the group heteroaryl-S— whereinthe heteroaryl group is as defined above including optionallysubstituted aryl groups as also defined above.

The term “thioether derivatives” when used to refer to the glycopeptidecompounds of this invention includes thioethers (—S—), sulfoxides (—SO—)and sulfones (—SO₂—).

As to any of the above groups which contain one or more substituents, itis understood, of course, that such groups do not contain anysubstitution or substitution patterns which are sterically impracticaland/or synthetically non-feasible. In addition, the compounds of thisinvention include all stereochemical isomers arising from thesubstitution of these compounds.

“Cyclodextrin” includes cyclic molecules containing six or moreα-D-glucopyranose units linked at the 1,4 positions by α linkages as inamylose. β-Cyclodextrin or cycloheptaamylose contains sevenα-D-glucopyranose units. As used herein, the term “cyclodextrin” alsoincludes cyclodextrin derivatives such as hydroxypropyl and sulfobutylether cyclodextrins. Such derivatives are described for example, in U.S.Pat. Nos. 4,727,064 and 5,376,645. One preferred cyclodextrin ishydroxypropyl β-cyclodextrin having a degree of substitution of fromabout 4.1-5.1 as measured by FTIR. Such a cyclodextrin is available fromCerestar (Hammond, Ind., USA) under the name Cavitron™ 82003.

“Glycopeptide” refers to oligopeptide (e.g. heptapeptide) antibiotics(dalbaheptides), characterized by a multi-ring peptide core optionallysubstituted with saccharide groups, such as vancomycin. Examples ofglycopeptides included in this definition may be found in “GlycopeptidesClassification, Occurrence, and Discovery”, by Raymond C. Rao and LouiseW. Crandall, (“Drugs and the Pharmaceutical Sciences” Volume 63, editedby Ramakrishnan Nagarajan, published by Marcal Dekker, Inc.). Additionalexamples of glycopeptides are disclosed in U.S. Pat. Nos. 4,639,433;4,643,987; 4,497,802; 4,698,327; 5,591,714; 5,840,684; and 5,843,889; inEP 0 802 199; EP 0 801 075; EP 0 667 353; WO 97/28812; WO 97/38702; WO98/52589; WO 98/52592; and in J. Amer. Chem. Soc., 1996, 118,13107-13108; J. Amer. Chem. Soc., 1997, 119, 12041-12047; and J. Amer.Chem. Soc., 1994, 116, 4573-4590. Representative glycopeptides includethose identified as A477, A35512, A40926, A41030, A42867, A47934,A80407, A82846, A83850, A84575, AB-65, Actaplanin, Actinoidin, Ardacin,Avoparcin, Azureomycin, Balhimycin, Chloroorienticin, Chloropolysporin,Decaplanin, N-demethylvancomycin, Eremomycin, Galacardin, Helvecardin,Izupeptin, Kibdelin, LL-AM374, Mannopeptin, MM45289, MM47756, MM47761,MM49721, MM47766, MM55260, MM55266, MM55270, MM56597, MM56598, OA-7653,Orenticin, Parvodicin, Ristocetin, Ristomycin, Symmonicin, Teicoplanin,UK-68597, UK-69542, UK-72051, Vancomycin, and the like. The term“glycopeptide” as used herein is also intended to include the generalclass of peptides disclosed above on which the sugar moiety is absent,i.e. the aglycone series of glycopeptides. For example, removal of thedisaccharide moiety appended to the phenol on vancomycin by mildhydrolysis gives vancomycin aglycone. Also within the scope of theinvention are glycopeptides that have been further appended withadditional saccharide residues, especially aminoglycosides, in a mannersimilar to vancosamine.

“Optional” or “optionally” means that the subsequently described eventor circumstance may or may not occur, and that the description includesinstances where said event or circumstance occurs and instances in whichit does not. For example, “optionally substituted” means that a groupmay or may not be substituted with the described substituent.

As used herein, the terms “inert organic solvent” or “inert solvent” or“inert diluent” mean a solvent or diluent which is essentially inertunder the conditions of the reaction in which it is employed as asolvent or diluent. Representative examples of materials which may beused as inert solvents or diluents include, by way of illustration,benzene, toluene, acetonitrile, tetrahydrofuran (“THF”),dimethylformamide (“DMF”), chloroform (“CHCl₃”), methylene chloride (ordichloromethane or “CH₂Cl₂), diethyl ether, ethyl acetate, acetone,methylethyl ketone, methanol, ethanol, propanol, isopropanol,tert-butanol, dioxane, pyridine, and the like. Unless specified to thecontrary, the solvents used in the reactions of the present inventionare inert solvents.

The term “nitrogen-linked” or “N-linked” means a group or substituent isattached to the remainder of a compound (e.g a compound of formula I)through a bond to a nitrogen of the group or substituent. The term“oxygen-linked” means a group or substituent is attached to theremainder of a compound (e.g. a compound of formula I) through a bond toan oxygen of the group or substituent. The term “sulfur-linked” means agroup or substituent is attached to the remainder of a compound (e.g. acompound of formula I) through a bond to a sulfur of the group orsubstituent.

“Pharmaceutically acceptable salt” means those salts which retain thebiological effectiveness and properties of the parent compounds andwhich are not biologically or otherwise harmful as the dosageadministered. The compounds of this invention are capable of formingboth acid and base salts by virtue of the presence of amino and carboxygroups respectively.

Pharmaceutically acceptable base addition salts may be prepared frominorganic and organic bases. Salts derived from inorganic bases include,but are not limited to, the sodium, potassium, lithium, ammonium,calcium, and magnesium salts. Salts derived from organic bases include,but are not limited to, salts of primary, secondary and tertiary amines,substituted amines including naturally-occurring substituted amines, andcyclic amines, including isopropylamine, trimethyl amine, diethylamine,triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol,tromethamine, lysine, arginine, histidine, caffeine, procaine,hydrabamine, choline, betaine, ethylenediamine, glucosamine,N-alkylglucamines, theobromine, purines, piperazine, piperidine, andN-ethylpiperidine. It should also be understood that other carboxylicacid derivatives would be useful in the practice of this invention, forexample carboxylic acid amides, including carboxamides, lower alkylcarboxamides, di(lower alkyl) carboxamides, and the like.

Pharmaceutically acceptable acid addition salts may be prepared frominorganic and organic acids. Salts derived from inorganic acids includehydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid and the like. Salts derived from organic acids includeacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid,malic acid, malonic acid, succinic acid, maleic acid, fumaric acid,tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid,salicylic acid and the like.

The compounds of this invention typically contain one or more chiralcenters. Accordingly, this invention is intended to include racemicmixtures, diasteromers, enantiomers and mixture enriched in one or moresteroisomer. The scope of the invention as described and claimedencompasses the racemic forms of the compounds as well as the individualenantiomers and non-racemic mixtures thereof.

The term “treatment” as used herein includes any treatment of acondition or disease in an animal, particularly a mammal, moreparticularly a human, and includes:

(i) preventing the disease or condition from occurring in a subjectwhich may be predisposed to the disease but has not yet been diagnosedas having it;

(ii) inhibiting the disease or condition, i.e. arresting itsdevelopment; relieving the disease or condition, i.e. causing regressionof the condition; or relieving the conditions caused by the disease,i.e. symptoms of the disease.

The term “disease state which is alleviated by treatment with a broadspectrum antibacterial” or “bacterial disease” as used herein isintended to cover all disease states which are generally acknowledged inthe art to be usefully treated with a broad spectrum antibacterial ingeneral, and those disease states which have been found to be usefullytreated by the specific antibacterials of this invention. Such diseasestates include, but are not limited to, treatment of a mammal afflictedwith pathogenic bacteria, in particular staphylococci (methicillinsensitive and resistant), streptococci (penicillin sensitive andresistant), enterococci (vancomycin sensitive and resistant), andClostridium difficile.

The term “therapeutically effective amount” refers to that amount whichis sufficient to effect treatment, as defined herein, when administeredto a mammal in need of such treatment. The therapeutically effectiveamount will vary depending on the subject and disease state beingtreated, the severity of the affliction and the manner ofadministration, and may be determined routinely by one of ordinary skillin the art.

The term “protecting group” or “blocking group” refers to any groupwhich, when bound to one or more hydroxyl, thiol, amino, carboxy orother groups of the compounds, prevents undesired reactions fromoccurring at these groups and which protecting group can be removed byconventional chemical or enzymatic steps to reestablish the hydroxyl,thio, amino, carboxy or other group. The particular removable blockinggroup employed is not critical and preferred removable hydroxyl blockinggroups include conventional substituents such as allyl, benzyl, acetyl,chloroacetyl, thiobenzyl, benzylidine, phenacyl, t-butyl-diphenylsilyland any other group that can be introduced chemically onto a hydroxylfunctionality and later selectively removed either by chemical orenzymatic methods in mild conditions compatible with the nature of theproduct. Protecting groups are disclosed in more detail in T. W. Greeneand P. G. M. Wuts, “Protective Groups in Organic Synthesis” 3^(rd) Ed.,1999, John Wiley and Sons, N.Y.

Preferred removable amino blocking groups include conventionalsubstituents such as t-butyoxycarbonyl (t-BOC), benzyloxycarbonyl (CBZ),fluorenylmethoxycarbonyl (FMOC), allyloxycarbonyl (ALOC) and the like,which can be removed by conventional conditions compatible with thenature of the product.

Preferred carboxy protecting groups include esters such as methyl,ethyl, propyl, t-butyl etc. which can be removed by mild conditionscompatible with the nature of the product.

“Vancomycin” refers to a glycopeptide antibiotic having the formula:

When describing vancomycin derivatives, the term “N^(van)—” indicatesthat a substituent is covalently attached to the amino group of thevacosamine moiety of vacomycin. Similarly, the term “N^(leu)—” indicatesthat a substituent is covalently attached to the amino group of theleucine moiety of vancomycin.

General Synthetic Procedures

The glycopeptide compounds of this invention can be prepared fromreadily available starting materials using the following general methodsand procedures. It will be appreciated that where typical or preferredprocess conditions (i.e., reaction temperatures, times, mole ratios ofreactants, solvents, pressures, etc.) are given, other processconditions can also be used unless otherwise stated. Optimum reactionconditions may vary with the particular reactants or solvent used, butsuch conditions can be determined by one skilled in the art by routineoptimization procedures.

Additionally, as will be apparent to those skilled in the art,conventional protecting groups may be necessary to prevent certainfunctional groups from undergoing undesired reactions. The choice of asuitable protecting group for a particular functional group as well assuitable conditions for protection and deprotection are well known inthe art. For example, numerous protecting groups, and their introductionand removal, are described in T. W. Greene and G. M. Wuts, ProtectingGroups in Organic Synthesis, Third Edition, Wiley, New York, 1999, andreferences cited therein.

In the following reaction schemes, the glycopeptide compounds aredepicted in a simplified form as a box “G” that shows the carboxyterminus labeled [C], the vancosamine amino terminus labeled [V], the“non-saccharide” amino terminus (leucine amine moiety) labeled [N], andoptionally, the resorcinol moiety labeled [R] as follows:

A glycopeptide compound of the present invention, which is substitutedat the C-terminus with a substituent that comprises one or more (e.g. 1,2, 3, 4, or 5) phosphono (—PO₃H) groups, can be prepared by coupling acorresponding glycopeptide compound wherein the C-terminus is a carboxygroup with a suitable phosphono containing compound. For example, aglycopeptide compound wherein the C-terminus is a carboxy group can becoupled with a phosphono containing amine, alcohol, or thiol compound toform an amide, an ester, or a thioester, respectively. For example aglycopeptide compound of formula I wherein R³ is a nitrogen linkedmoiety comprising one or more phosphono groups can be prepared bycoupling a corresponding glycopeptide compound of formula I wherein R³is hydroxy with the requisite phosphono-containing amine to form theformula I wherein R³ is a nitrogen linked moiety comprising one or morephosphono groups.

A glycopeptide compound of the present invention, which is substitutedat the C-terminus with a substituent that comprises one or more (e.g. 1,2, 3, 4, or 5) phosphono (—PO₃H₂) groups, and wherein the vancosamineamino terminus (V) is substituted, can be prepared by first reductivelyalkylating the corresponding glycopeptide compound wherein thevancosamine amino terminus (V) is the free amine (NH₂) and then couplingthe corresponding glycopeptide compound with the requisite phosphonocontaining compound (e.g. phosphono containing amine, alcohol, orthiol).

By way of illustration, a glycopeptide compound, such as vancomycin, canfirst be reductive alkylated as shown in the following reaction:

where A represents R^(a) minus one carbon atom and R^(a), R^(b), Y, Zand x are as defined herein. This reaction is typically conducted byfirst contacting one equivalent of the glycopeptide, i.e., vancomycin,with an excess, preferably from 1.1 to 1.3 equivalents, of the desiredaldehyde in the presence of an excess, preferably about 2.0 equivalents,of a tertiary amine, such as diisopropylethylamine (DIPEA) and the like.This reaction is typically conducted in an inert diluent, such as DMF oracetonitrile/water, at ambient temperature for about 0.25 to 2 hoursuntil formation of the corresponding imine and/or hemiaminal issubstantially complete. The resulting imine and/or hemiaminal istypically not isolated, but is reacted in situ with a reducing agent,such as sodium cyanoborohydride, pyridine borane, or the like, to affordthe corresponding amine. This reaction is preferably conducted bycontacting the imine and/or hemiaminal with an excess, preferably about3 equivalents, of trifluoroacetic acid, followed by about 1 to 12equivalents of the reducing agent at ambient temperature in methanol oracetonitrile/water. The resulting alkylated product is readily purifiedby conventional procedures, such as precipitation and/or reverse-phaseHPLC. Surprisingly, by forming the imine and/or hemiaminal in thepresence of a trialkyl amine, and then acidifying with trifluoroaceticacid before contact with the reducing agent, the selectivity for thereductive alkylating reaction is greatly improved, i.e., reductivealkylating at the amino group of the saccharide (e.g., vancosamine) isfavored over reductive alkylating at the N-terminus (e.g., the leucinylgroup) by at least 10:1, more preferably 20:1.

The above process is a significantly improvement over previous methodsfor selectively alkylating an amino saccharide group of a glycopeptideantibiotic. Thus, the present invention also provides a method foralkylating a glycopeptide that comprises a saccharide-amine comprising:

combining an aldehyde or ketone, a suitable base, and the glycopeptide,to provide a reaction mixture;

acidifying the reaction mixture; and

combining the reaction mixture with a suitable reducing agent, toprovide a glycopeptide that is alkylated at the saccharide-amine.Preferably, the glycopeptide comprises at least one amino group otherthan the saccharide-amine.

Preferably, the reductive alkylating at the saccharide-amine is favoredover reductive alkylating at another amino group of the glycopeptide byat least about 10:1; and more preferably, by at least about 15:1 orabout 20:1.

The reductive alkylating process of the invention is typically carriedout in the presence of a suitable solvent or combination of solvents,such as, for example, a halogenated hydrocarbon (e.g. methylenechloride), a linear or branched ether (e.g. diethyl ether,tetrahydrofuran), an aromatic hydrocarbon (e.g. benzene or toluene), analcohol (methanol, ethanol, or isopropanol), dimethylsulfoxide (DMSO),N,N-dimethylformamide, acetonitrile, water,1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidone, tetramethyl urea,N,N-dimethylacetamide, diethylformamide (DMF), 1-methyl-2-pyrrolidinone,tetramethylenesulfoxide, glycerol, ethyl acetate, isopropyl acetate,N,N-dimethylpropylene urea (DMPU) or dioxane. Preferably the alkylatingis carried out in acetonitrile/water, or DMF/methanol.

Preferably the reduction (i.e. treatment with the reducing agent) iscarried out in the presence of a protic solvent, such as, for example,an alcohol (e.g. methanol, ethanol, propanol, isopropanol, or butanol),water, or the like.

The reductive alkylating process of the invention can be carried out atany suitable temperature from the freezing point to the refluxtemperature of the reaction mixture. Preferably the reaction is carriedout at a temperature in the range of about 0° C. to about 100° C. Morepreferably at a temperature in a range of about 0° C. to about 50° C.,or in a range of about 20° C. to about 30° C.

Any suitable base can be employed in the reductive alkylating process ofthe invention. Suitable bases include tertiary amines (e.g.diisopropylethylamine, N-methylmorpholine or triethylamine) and thelike.

Any suitable acid can be used to acidify the reaction mixture. Suitableacids include carboxylic acids (e.g. acetic acid, trichloroacetic acid,citric acid, formic acid, or trifluoroacetic acid), mineral acids (e.g.hydrochloric acid, sulfuric acid, or phosphoric acid), and the like. Apreferred acid is trifluoroacetic acid.

Suitable reducing agents for carrying out reductive alkylating processof the invention are known in the art. Any suitable reducing agent canbe employed in the methods of the invention, provided it is compatiblewith the functionality present in the glycopeptide. For example,suitable reducing agents include sodium cyanoborohydride, sodiumtriacetoxyborohydride, pyridine/borane, sodium borohydride, and zincborohydride. The reduction can also be carried out in the presence of atransition metal catalyst (e.g. palladium or platinum) in the presenceof a hydrogen source (e.g. hydrogen gas or cyclohexadiene). See forexample, Advanced Organic Chemistry, Fourth Edition, John Wiley & Sons,New York (1992), 899-900.

The glycopeptide derivative resulting from the reductive alkylating isthen coupled with a phosphono containing amine (R³—H) to form an amidebond. This reaction is illustrated by the following reaction:

where R³ is a nitrogen-linked group that comprises one or more phosphonogroups. In this reaction, the glycopeptide derivative is typicallycontacted with the amine in the presence of a peptide coupling reagent,such as PyBOP and HOBT, to provide the amide. This reaction is typicallyconducted in an inert diluent, such as DMF, at a temperature rangingfrom about 0° C. to about 60° C. for about 1 to 24 hours or until thecoupling reaction is substantially complete. Subsequent deprotectionusing conventional procedures and reagents affords the compound of thisinvention.

If desired, the amine coupling step described above can be conductedfirst to provide an amide, followed by reductive alkylating anddeprotection to afford the compound of the invention.

If desired, the glycopeptide compounds of this invention can also beprepared in a step-wise manner in which a precursor to the—R^(a)—Y—R^(b)—(Z) group is first attached the glycopeptide by reductivealkylating, followed by subsequent elaboration of the attached precursorusing conventional reagent and procedures to form the—R^(a)—Y—R^(b)—(Z)_(x) group. Additionally, ketones may also be employedin the above-described reductive alkylating reactions to affordα-substituted amines.

Any glycopeptide having an amino group may be employed in thesereductive alkylating reactions. Such glycopeptides are well-known in theart and are either commercially available or may be isolated usingconventional procedures. Suitable glycopeptides are disclosed, by way ofexample, in U.S. Pat. Nos. 3,067,099; 3,338,786; 3,803,306; 3,928,571;3,952,095; 4,029,769; 4,051,237; 4,064,233; 4,122,168; 4,239,751;4,303,646; 4,322,343; 4,378,348; 4,497,802; 4,504,467; 4,542,018;4,547,488; 4,548,925; 4,548,974; 4,552,701; 4,558,008; 4,639,433;4,643,987; 4,661,470; 4,694,069; 4,698,327; 4,782,042; 4,914,187;4,935,238; 4,946,941; 4,994,555; 4,996,148; 5,187,082; 5,192,742;5,312,738; 5,451,570; 5,591,714; 5,721,208; 5,750,509; 5,840,684; and5,843,889. Preferably, the glycopeptide employed in the above reactionis vancomycin.

As illustrated in the following scheme, a phosphono containingaminoalkyl sidechain at the resorcinol moiety of a glycopeptide, such asvancomycin, can be introduced via a Mannich reaction (in this scheme,the resorcinol moiety of the glycopeptide is illustrated for clarity).In this reaction, an amine of formula NHRR′ (wherein one or both of Rand R′ is a group that comprises one or more phosphono groups), and analdehyde (e.g. CH₂O), such as formalin (a source of formaldehyde), arereacted with the glycopeptide under basic conditions to give theglycopeptide derivative.

Compounds of the invention comprising a sulfoxide or sulfone can beprepared from the corresponding thio compounds using conventionalreagents and procedures. Suitable reagents for oxidizing a thio compoundto a sulfoxide include, by way of example, hydrogen peroxide, peracidessuch as 3-chloroperoxybenzoic acid (MCPBA), sodium periodate, sodiumchlorite, sodium hypochlorite, calcium hypochlorite, tert-butylhypochlorite and the like. Chiral oxidizing reagents, (optically activereagents) may also be employed to provide chiral sulfoxides. Suchoptically active reagents are well-known in the art and include, forexample, the reagents described in Kagen et al., Synlett., 1990,643-650.

The aldehydes and ketones employed in the above reactive alkylatingreactions are also well-known in the art and are either commerciallyavailable or can be prepared by conventional procedures usingcommercially available starting materials and conventional reagents (forexample see March, Advanced Organic Chemistry, Fourth Edition, JohnWiley & Sons, New York (1992), and references cited therein).

The phosphono substituted compounds (e.g. the phosphono substitutedamines, alcohols, or thiols) are either commercially available or can beprepared by conventional procedures using commercially availablestarting materials and reagents. See for example, Advanced OrganicChemistry, Jerry March, 4th ed., 1992, John Wiley and Sons, New York,page 959; and Frank R. Hartley (ed.) The Chemistry of OrganophosphorousCompounds, vol. 1-4, John Wiley and Sons, New York (1996).Aminomethylphosphonic acid is commercially available from AldrichChemical Company, Milwaukee, Wis.

Additional details and other methods for preparing the compounds of thisinvention are described in the Examples below.

Pharmaceutical Compositions

This invention also includes pharmaceutical composition containing thenovel glycopeptide compounds of this invention. Accordingly, theglycopeptide compound, preferably in the form of a pharmaceuticallyacceptable salt, can be formulated for oral or parenteral administrationfor the therapeutic or prophylactic treatment of bacterial infections.

By way of illustration, the glycopeptide compound can be admixed withconventional pharmaceutical carriers and excipients and used in the formof tablets, capsules, elixirs, suspensions, syrups, wafers, and thelike. Such pharmaceutical compositions will contain from about 0.1 toabout 90% by weight of the active compound, and more generally fromabout 10 to about 30%. The pharmaceutical compositions may containcommon carriers and excipients, such as corn starch or gelatin, lactose,sucrose, microcrystalline cellulose, kaolin, mannitol, dicalciumphosphate, sodium chloride, and alginic acid. Disintegrators commonlyused in the formulations of this invention include croscarmellose,microcrystalline cellulose, corn starch, sodium starch glycolate andalginic acid.

A liquid composition will generally consist of a suspension or solutionof the compound or pharmaceutically acceptable salt in a suitable liquidcarrier(s), for example ethanol, glycerine, sorbitol, non-aqueoussolvent such as polyethylene glycol, oils or water, optionally with asuspending agent, a solubilizing agent (such as a cyclodextrin),preservative, surfactant, wetting agent, flavoring or coloring agent.Alternatively, a liquid formulation can be prepared from areconstitutable powder.

For example a powder containing active compound, suspending agent,sucrose and a sweetener can be reconstituted with water to form asuspension; and a syrup can be prepared from a powder containing activeingredient, sucrose and a sweetener.

A composition in the form of a tablet can be prepared using any suitablepharmaceutical carrier(s) routinely used for preparing solidcompositions. Examples of such carriers include magnesium stearate,starch, lactose, sucrose, microcrystalline cellulose and binders, forexample polyvinylpyrrolidone. The tablet can also be provided with acolor film coating, or color included as part of the carrier(s). Inaddition, active compound can be formulated in a controlled releasedosage form as a tablet comprising a hydrophilic or hydrophobic matrix.

A composition in the form of a capsule can be prepared using routineencapsulation procedures, for example by incorporation of activecompound and excipients into a hard gelatin capsule. Alternatively, asemi-solid matrix of active compound and high molecular weightpolyethylene glycol can be prepared and filled into a hard gelatincapsule; or a solution of active compound in polyethylene glycol or asuspension in edible oil, for example liquid paraffin or fractionatedcoconut oil can be prepared and filled into a soft gelatin capsule.

Tablet binders that can be included are acacia, methylcellulose, sodiumcarboxymethylcellulose, poly-vinylpyrrolidone (Povidone), hydroxypropylmethylcellulose, sucrose, starch and ethylcellulose. Lubricants that canbe used include magnesium stearate or other metallic stearates, stearicacid, silicone fluid, talc, waxes, oils and colloidal silica.

Flavoring agents such as peppermint, oil of wintergreen, cherryflavoring or the like can also be used. Additionally, it may bedesirable to add a coloring agent to make the dosage form moreattractive in appearance or to help identify the product.

The compounds of the invention and their pharmaceutically acceptablesalts that are active when given parenterally can be formulated forintramuscular, intrathecal, or intravenous administration.

A typical composition for intra-muscular or intrathecal administrationwill consist of a suspension or solution of active ingredient in an oil,for example arachis oil or sesame oil. A typical composition forintravenous or intrathecal administration will consist of a sterileisotonic aqueous solution containing, for example active ingredient anddextrose or sodium chloride, or a mixture of dextrose and sodiumchloride. Other examples are lactated Ringer's injection, lactatedRinger's plus dextrose injection, Normosol-M and dextrose, Isolyte E,acylated Ringer's injection, and the like. Optionally, a co-solvent, forexample, polyethylene glycol; a chelating agent, for example,ethylenediamine tetracetic acid; a solubilizing agent, for example, acyclodextrin; and an anti-oxidant, for example, sodium metabisulphite,may be included in the formulation. Alternatively, the solution can befreeze dried and then reconstituted with a suitable solvent just priorto administration.

In a preferred embodiment, the glycopeptide derivatives of thisinvention are formulated in an aqueous solution containing acyclodextrin. In another preferred embodiment the glycopeptidederivatives of this invention are formulated as a lyophilized powdercontaining a cyclodextrin or as a sterile powder containing acyclodextrin. Preferably, the cyclodextrin ishydroxypropyl-β-cyclodextrin or sulfobutyl ether β-cyclodextrin; morepreferably, the cyclodextrin is hydroxypropyl-β-cyclodextrin. Typically,in an injectable solution, the cyclodextrin will comprise about 1 to 25weight percent; preferably, about 2 to 10 weight percent; morepreferable, about 4 to 6 weight percent, of the formulation.Additionally, the weight ratio of the cyclodextrin to the glycopeptidederivative will preferably be from about 1:1 to about 10:1.

The compounds of the invention and their pharmaceutically acceptablesalts which are active on rectal administration can be formulated assuppositories. A typical suppository formulation will generally consistof active ingredient with a binding and/or lubricating agent such as agelatin or cocoa butter or other low melting vegetable or synthetic waxor fat.

The compounds of this invention and their pharmaceutically acceptablesalts which are active on topical administration can be formulated astransdermal compositions or transdermal delivery devices (“patches”).Such compositions include, for example, a backing, active compoundreservoir, a control membrane, liner and contact adhesive. Suchtransdermal patches may be used to provide continuous or discontinuousinfusion of the compounds of the present invention in controlledamounts. The construction and use of transdermal patches for thedelivery of pharmaceutical agents is well known in the art. See, e.g.,U.S. Pat. No. 5,023,252, issued Jun. 11, 1991. Such patches may beconstructed for continuous, pulsatile, or on demand delivery ofpharmaceutical agents.

The active compound is effective over a wide dosage range and isgenerally administered in a pharmaceutically effective amount. It, willbe understood, however, that the amount of the compound actuallyadministered will be determined by a physician, in the light of therelevant circumstances, including the condition to be treated, thechosen route of administration, the actual compound administered and itsrelative activity, the age, weight, and response of the individualpatient, the severity of the patient's symptoms, and the like.

Suitable doses are in the general range of from 0.01-100 mg/kg/day,preferably 0.1-50 mg/kg/day. For an average 70 kg human, this wouldamount to 0.7 mg to 7 g per day, or preferably 7 mg to 3.5 g per day. Amore preferred dose for a human is about 500 mg to about 2 g per day.

Other suitable formulations for use in the present invention can befound in Remington's Pharmaceutical Sciences, Mace Publishing Company,Philadelphia, Pa., 17th ed. (1985).

The following illustrate representative pharmaceutical compositions ofthe present invention.

FORMULATION EXAMPLE A

This example illustrates the preparation of a representativepharmaceutical composition for oral administration of a compound of thisinvention:

Ingredients Quantity per tablet, (mg) Active Compound 200 Lactose,spray-dried 148 Magnesium stearate 2The above ingredients are mixed and introduced into a hard-shell gelatincapsule.

FORMULATION EXAMPLE B

This example illustrates the preparation of another representativepharmaceutical composition for oral administration of a compound of thisinvention:

Ingredients Quantity per tablet, (mg) Active Compound 400 Cornstarch 50Lactose 145 Magnesium stearate 5The above ingredients are mixed intimately and pressed into singlescored tablets.

FORMULATION EXAMPLE C

This example illustrates the preparation of a representativepharmaceutical composition for oral administration of a compound of thisinvention.

An oral suspension is prepared having the following composition.

Ingredients Active Compound 1.0 g Fumaric acid 0.5 g Sodium chloride 2.0g Methyl paraben 0.1 g Granulated sugar 25.5 g Sorbitol (70% solution)12.85 g Veegum K (Vanderbilt Co.) 1.0 g Flavoring 0.035 ml Colorings 0.5mg Distilled water q.s. to 100 ml

FORMULATION EXAMPLE D

This example illustrates the preparation of a representativepharmaceutical composition containing a compound of this invention.

An injectable preparation buffered to a pH of 4 is prepared having thefollowing composition:

Ingredients Active Compound 0.2 g Sodium Acetate Buffer Solution (0.4M)2.0 ml HCl (1N) q.s. to pH 4 Water (distilled, sterile) q.s. to 20 ml

FORMULATION EXAMPLE E

This example illustrates the preparation of a representativepharmaceutical composition for injection of a compound of thisinvention.

A reconstituted solution is prepared by adding 20 ml of sterile water to1 g of the compound of this invention. Before use, the solution is thendiluted with 200 ml of an intravenous fluid that is compatible with theactive compound. Such fluids are chosen from 5% dextrose solution, 0.9%sodium chloride, or a mixture of 5% dextrose and 0.9% sodium chloride.Other examples are lactated Ringer's injection, lactated Ringer's plus5% dextrose injection, Normosol-M and 5% dextrose, Isolyte E, andacylated Ringer's injection

FORMULATION EXAMPLE F

This example illustrates the preparation of a representativepharmaceutical composition containing a compound of this invention.

An injectable preparation is prepared having the following composition:

Ingredients Active Compound 0.1-5.0 g Hydroxypropyl-β-cyclodextrin 1-25g 5% Aqueous Dextrose Solution (sterile) q.s. to 100 mlThe above ingredients are blended and the pH is adjusted to 3.5±0.5using 0.5 N HCl or 0.5 N NaOH.

FORMULATION EXAMPLE G

This example illustrates the preparation of a representativepharmaceutical composition containing a compound of this invention.

A frozen solution suitable for injection is prepared having thefollowing composition:

Frozen Solution Active Compound 250 mg to 1000 mgHydroxypropyl-β-cyclodextrin 250 mg to 10 g Excipients - e.g., dextrose0-50 g Water for Injection 10-100 ml The weight ratio ofhydroxypropyl-β-cyclodextrin to the active compound will typically befrom about 1:1 to about 10:1.

-   -   Representative Procedure: Hydroxypropyl-β-cyclodextrin and        excipients, if any, are dissolved in about 80% of the water for        injection and the active compound is added and dissolved. The pH        is adjusted with 1 M sodium hydroxide to 4.7±0.3 and the volume        is then adjusted to 95% of the final volume with water for        injection. The pH is checked and adjusted, if necessary, and the        volume is adjusted to the final volume with water for injection.        The formulation is then sterile filtered through a 0.22 micron        filter and placed into a sterile vial under aseptic conditions.        The vial is capped, labeled and stored frozen.

FORMULATION EXAMPLE H

This example illustrates the preparation of a representativepharmaceutical composition containing a compound of this invention.

A lyophilized powder useful for preparing an injectable solution isprepared having the following composition:

Lyophilized Powder Active Compound 250 mg to 1000 mgHydroxypropyl-β-cyclodextrin 250 mg to 10 g Excipients - e.g., mannitol,0-50 g sucrose and/or lactose Buffer agent - e.g., citrate 0-500 mg Theweight ratio of hydroxypropyl-β-cyclodextrin to the active compound willtypically be from about 1:1 to about 10:1.

-   -   Representative Procedure: Hydroxypropyl-β-cyclodextrin and        excipients and/or buffering agents, if any, are dissolved in        about 60% of the water for injection. The active compound is        added and dissolved and the pH is adjusted with 1 M sodium        hydroxide to 4.0-5.0 and the volume is adjusted to 95% of the        final volume with water for injection. The pH is checked and        adjusted, if necessary, and the volume is adjusted to the final        volume with water for injection. The formulation is then sterile        filtered through a 0.22 micron filter and placed into a sterile        vial under aseptic conditions. The formulation is then        freeze-dried using an appropriate lyophilization cycle. The vial        is capped (optionally under partial vacuum or dry nitrogen),        labeled and stored at room temperature or under refrigeration.

FORMULATION EXAMPLE I

This example illustrates the preparation of a representativepharmaceutical composition containing a compound of this invention.

A sterile powder useful for preparing an injectable solution is preparedhaving the following composition:

Sterile Powder Active Compound 250 mg to 1000 mgHydroxypropyl-β-cyclodextrin 250 mg to 10 g¹ Excipients optional Theweight ratio of hydroxypropyl-β-cyclodextrin to the active willtypically be from about 1:1 to about 10:1.

-   -   Representative Procedure: Hydroxypropyl-β-cyclodextrin and the        active compound (and any excipients) are dispersed into an        appropriate sterile container and the container is sealed        (optionally under partial vacuum or dry nitrogen), labeled and        stored at room temperature or under refrigeration.        Administration of Representative Formulations H and I to a        Patient

The pharmaceutical formulations described in formulation examples H andI above can be administered intravenously to a patient by theappropriate medical personnel to treat or prevent gram-positiveinfections. For administration, the above formulations can bereconstituted and/or diluted with a diluent, such as 5% dextrose orsterile saline, as follows:

-   -   Representative Procedure: The lyophilized powder of formulation        example H (e.g., containing 1000 mg of active compound) is        reconstituted with 20 ml of sterile water and the resulting        solution is further diluted with 80 ml of sterile saline in a        100 ml infusion bag. The diluted solution is then administered        to the patient intravenously over 30 to 120 minutes.

FORMULATION EXAMPLE J

This example illustrates the preparation of a representativepharmaceutical composition for topical application of a compound of thisinvention.

Ingredients grams Active compound 0.2-10 Span 60 2 Tween 60 2 Mineraloil 5 Petrolatum 10 Methyl paraben 0.15 Propyl paraben 0.05 BHA(butylated hydroxy anisole) 0.01 Water q.s. to 100All of the above ingredients, except water, are combined and heated to60° C. with stirring. A sufficient quantity of water at 60° C. is thenadded with vigorous stirring to emulsify the ingredients, and water thenadded q.s. 100 g.

FORMULATION EXAMPLE K

This example illustrates the preparation of a representativepharmaceutical composition containing a compound of this invention.

A suppository totaling 2.5 grams is prepared having the followingcomposition:

Ingredients Active Compound 500 mg Witepsol H-15* balance(*triglycerides of saturated vegetable fatty acid; a product ofRiches-Nelson, Inc., New York, N.Y.)

A preferred active compound for incorporation in Formulations A-K iscompound II, or a pharmaceutically acceptable salt thereof (e.g. thehydrochloride salt).

Utility

The glycopeptide compounds of this invention, and their pharmaceuticallyacceptable salts, are useful in medical treatments and exhibitbiological activity, including antibacterial activity, which can bedemonstrated in using the tests described herein. Such tests are wellknown to those skilled in the art, and are referenced and described inLorian “Antibiotics in Laboratory Medicine”, Fourth Edition, Williamsand Wilkins (1991).

Accordingly, this invention provides methods for treating bacterial orinfectious diseases, especially those caused by Gram-positivemicroorganisms, in animals. The compounds of this invention areparticularly useful in treating infections caused bymethicillin-resistant staphylococci. Also, the compounds are useful intreating infection due to enterococci, including vancomycin-resistantenterococci (VRE). Examples of such diseases include severestaphylococcal infections, such as staphylococcal endocarditis andstaphylococcal septicemia. The animal treated may be either susceptibleto, or infected with, the microorganism. The method of treatmenttypically comprises administering to the animal an amount of a compoundof this invention which is effective for this purpose.

In practicing this method, the antibiotic can be administered in asingle daily dose or in multiple doses per day. The treatment regimenmay require administration over extended periods of time, for example,for several days or for from one to six weeks. The amount peradministered dose or the total amount administered will depend on suchfactors as the nature and severity of the infection, the age and generalhealth of the patient, the tolerance of the patient to the antibioticand the microorganism or microorganisms in the infection. Preferably,the compounds of the invention are administered intervenously.

Among other properties, the glycopeptide compounds of the invention havebeen found to have reduced mammalian toxicity when administered to amammal. For example, the phosphono substituted derivatives of theinvention have been found to have reduced liver and/or kidneyaccumulation compared to the corresponding non-phosphono substitutedcompounds. Moreover, certain compounds of this invention are expected tohave reduced nephrotoxicity. Additionally, it has been discovered thatthe addition of a cyclodextrin compound to a pharmaceutical compositioncontaining the glycopeptide compounds of this invention further reducesthe nephrotoxicity and/or tissue accumulation of the glycopeptidecompound when administered to a mammal.

The following synthetic and biological examples are offered toillustrate this invention and are not to be construed in any way aslimiting the scope of this invention.

EXAMPLES

In the examples below, the following abbreviations have the followingmeanings. Any abbreviations not defined have their generally acceptedmeaning. Unless otherwise stated, all temperatures are in degreesCelsius.

ACN = acetonitrile BOC, Boc = tert-butoxycarbonyl DIBAL-H =diisobutylaluminum hydride DIPEA = diisopropylethylamine DMF =N,N-dimethylformamide DMSO = dimethyl sulfoxide eq. = equivalent EtOAc =ethyl acetate Fmoc = 9-fluorenylmethoxycarbonyl HOBT =1-hydroxybenzotriazole hydrate Me = methyl MS = mass spectroscopy PyBOP= benzotriazol-1-yloxytris(pyrrolidino)phosphonium hexafluorophosphateTEMPO = 2,2,6,6-tetramethyl-piperidinyloxy, free radical TFA =trifluoroacetic acid THF = tetrahydrofuran TLC, tlc = thin layerchromatography

In the following examples, vancomycin hydrochloride semi-hydrate waspurchased from Alpharma, Inc. Fort Lee, N.J. 07024 (Alpharma AS, OsloNorway). Other reagents and reactants are available from AldrichChemical Co., Milwaukee, Wis. 53201.

General Procedure A Reductive Alkylating of Vancomycin

To a mixture of vancomycin (1 eq.) and the desired aldehyde (1.3 eq.) inDMF was added DIPEA (2 eq.). The reaction was stirred at ambienttemperature for 1-2 hours and monitored by reverse-phase HPLC. Methanoland NaCNBH₃ (1 eq.) were added to the solution, followed by TFA (3 eq.).Stirring was continued for an additional hour at ambient temperature.After the reaction was complete, the methanol was removed in vacuo. Theresidue was precipitated in acetonitrile. Filtration gave the crudeproduct which was then purified by reverse-phase HPLC. If desired, otherglycopeptides antibiotics may be used in this procedure.

General Procedure B Synthesis of 2-(Decylthio)acetaldehyde

Under nitrogen, to a suspension of potassium carbonate (27 g, 200 mmol)in acetone (100 ml) was added decyl bromide (10 ml, 50 mmol) andmercaptoethanol (4.4 ml, 63 mmol). The suspension was stirred at roomtemperature for 2 days, then partitioned between water and 80%hexane/ethyl acetate. The organic phase was washed with 2N sodiumhydroxide, dried over magnesium sulfate, and the volatiles removed undervacuum to give 2-(decylthio)ethanol (10.2 g, 47 mmol) as a colorlessliquid that was used without further purification.

Under nitrogen, 2-decylthioethanol (50 g, 230 mmol),N,N-diisopropylethylamine (128 ml, 730 mmol) and methylene chloride (400ml) were cooled to −40° C. To this solution was added, over 15 minutes,a solution of sulfur trioxide pyridine complex (116 g, 730 mmol) indimethyl sulfoxide (600 ml) and methylene chloride (200 ml). Afteraddition, the mixture was stirred a further 15 minutes at 40° C., then600 ml ice water as added. The mixture was removed from the coolingbath, 1 L water was added, and the liquids partitioned. The organicphase was washed with 1 L of 1 N hydrochloric acid, and dried overmagnesium sulfate. Filtration gave 600 ml liquid, which was diluted with600 ml hexane and passed through 200 ml silica. The silica was washedwith 100 ml 50% methylene chloride/hexane, then 300 ml methylenechloride. The combined organics were concentrated in vacuo to give2-(decylthio)acetaldehyde (48 g, 220 mmol) as a colorless liquid thatwas used without further purification.

General Procedure C Synthesis of N^(van)-2-(Decylthio)ethyl Vancomycin

Procedure A: Under nitrogen, vancomycin hydrochloride hydrate (1 g, 0.64mmol) was added to 2-(decylthio)acetaldehyde (139 mg, 0.64 mmol) inN,N-dimethylformamide (8 ml). N,N-diisopropylethylamine (336 uL, 1.9mmol) was added and the suspension stirred vigorously for 2.5 hours,over the course of which all the vancomycin dissolved. Solid sodiumcyanoborohydride (60 mg, 0.96 mmol) was added, followed by methanol (5ml) and trifluoroacetic acid (250 uL, 3.2 mmol). The reaction wasstirred for 55 minutes at room temperature and analyzed by reverse phaseHPLC. The product distribution based on uv absorption at 280 nm was asfollows:

Elution time (min) Area % Product 2.0 29 vancomycin 3.1 50N^(van)-2-(decylthio)ethyl vancomycin 3.2 2 — 3.3 7N^(leu)-2-(decylthio)ethyl vancomycin 3.9 13 N^(van),N^(leu)-bis-[2-(decylthio)ethyl] vancomycin 4.0 0.5 —

Procedure B: Under nitrogen, to a solution of 2-(decylthio)acetaldehyde(crude, 48 g, 220 mmol) in N,N-dimethylformamide (1.4 L) was added solidvancomycin hydrochloride hydrate (173 g, 110 mmol) followed byN,N-diisopropylethylamine (58 ml, 330 mmol). The suspension was stirredvigorously at room temperature for 2 hours, in the course of which timeall the vancomycin fully dissolved, then trifluoroacetic acid (53 ml,690 mmol) was added. The solution was stirred a further 90 minutes, thensolid sodium cyanoborohydride (10.5 g, 170 mmol) followed by methanol(800 ml) were added. After three hours the reaction was analyzed byreverse-phase HPLC. The product distribution based on uv absorption at280 nm was as follows:

Elution time (min) Area % Product 2.0 15 vancomycin 3.2 77N^(van)-2-(decylthio)ethyl vancomycin 3.3 3 — 3.4 0.5N^(leu)-2-(decylthio)ethyl vancomycin 4.0 0.8 N^(van),N^(leu)-bis-[2-(decylthio)ethyl] vancomycin 4.1 4 —

The reaction mixture from either of the above procedures was poured intowater (7 L), resulting in a slightly cloudy solution. The pH of thesolution was adjusted to 5 with saturated sodium bicarbonate, resultingin the formation of a white precipitate. This precipitate was collectedby filtration, washed with water then ethyl acetate and dried undervacuum to afford N^(van)-2-(decylthio)ethyl vancomycin, which was usedwithout further purification.

Procedure C: A solution of vancomycin hydrochloride (3.0 g, 2.1 mmol) inACN/H₂O (1:1, 30 ml) was treated with diisopropylethylamine (0.54 g,0.72 ml, 4.2 mmol) followed by 2-(decylthio)acetaldehyde (0.91 g, 4.2mmol) at 25° C. After 30 min the reaction mixture was treated with TFA(1.92 g, 1.29 ml, 16.8 mmol) followed by NaCNBH₃ (0.132 g, 2.1 mmol).After 5 to 10 minutes, the crude product N′-2-(decylthio)ethylvancomycin is precipitated in acetonitrile (300 ml).

Example 1 Preparation of Compound 3 Formula II wherein R³ isN-(phosphonomethyl)-amino; R⁵ is hydrogen; R¹⁹ is hydrogen, and R²⁰ is—CH₂CH₂—S—(CH₂)₉CH₃

N^(VAN)-(2-decylthio)ethyl vancomycin bistrifluoroacetate (1 g, 0.53mmol) and diisopropylethylamine (0.23 ml, 1.33 mmol) were combined inDMF (10 ml) and stirred until homogeneous. HOBt (0.080 g, 0.58 mmol) andPYBOP (0.300 g, 0.58 mmol) were then added to the reaction mixture.After 5-10 minutes a homogeneous solution containing(aminomethyl)phosphonic acid (0.060 g, 0.53 mmol) anddiisopropylethylamine (0.23 ml, 1.33 mmol) in water (3 ml) was added.The reaction was stirred at room temperature and monitored by MS. Whenthe reaction was judged to be complete, the reaction mixture was dilutedwith acetonitrile (40 ml) and centrifuged. The supernatant was discardedand the remaining pellet containing desired product was dissolved in 50%aqueous acetonitrile (10 ml) and purified by reverse phase preparativeHPLC to give the title compound. MS calculated (M+) 1742.7; found (MH+)1743.6.

Example 2 Preparation of Compound 11 Formula II wherein R³ is —OH; R⁵N-phosphonomethyl)-aminomethyl; R¹⁹ is hydrogen, and R²⁰ is—CH₂CH₂—NH—(CH₂)₉CH₃

(Aminomethyl)phosphonic acid (3.88 g, 35 mmol) and diisopropylethylamine(6.1 ml, 35 mmol) were combined in water (40 ml) and stirred untilhomogeneous. Acetonitrile (50 ml) and formaldehyde (37% solution in H₂O;0.42 ml, 05.6 mmol) were then added to the reaction mixture. Afterapproximately 15 minutes both N^(VAN)-decylaminoethyl vancomycintristrifluoroacetate (10.0 g, 5.1 mmol) and diisopropylethylamine (6.1ml, 35 mmol) were added to the reaction mixture. The reaction wasstirred at room temperature for approximately 18 hrs, at which time thepH was adjusted to about 7 with 20% TFA, acetonitrile was removed invacuo, and the residue was lyophylized. The resulting solid wastriturated with water (100 mL), collected by filtration, dried in vacuoand purified by reverse phase preparative HPLC to give the titlecompound. MS calculated (MH+) 1756.6; found (MH+) 1756.6.

Compound 11 was also prepared as follows.

The quinuclidine salt of N^(VAN)-(decylaminoethyl)vancomycin (500 mg,0.28 mmol, sub-part f below) and aminomethylphosphonic acid (155 mg, 1.4mmol) were slurried in 50% aqueous acetonitrile (10 mL).Diisopropyl-ethylamine (972 uL, 720 mg, 5.6 mmol) was added and themixture stirred at room temperature until the solids had dissolved. Thereaction mixture was then cooled in an ice bath and formalin (3.7%, madeby diluting commercial 37% formalin 1:9 with 50% ACN/water, 220 uL, 8.8mg, 0.29 mmol) was added. The reaction mixture was stirred at 0° for 15hours, at which time the reaction to be complete. The reaction wasquenched at 0° by adding 3N HCl to about pH 2. The mixture was dilutedto 50 mL with 50% ACN/water, and then acetonitrile was added (75 mL,followed by 5×10 mL at 5 minute intervals, 125 mL total) to precipitatethe product. The solid was collected by vacuum filtration and dried invacuo. Purification by reverse phase preparative HPLC gave the titlecompound.

The intermediate N^(VAN)-decylaminoethyl vancomycin tristrifluoroacetatewas prepared as follows.

-   a. N-Fmoc-2-(decylamino)ethanol. 2-(n-Decylamino)ethanol (2.3 g, 111    mmol, 1.1 eq) and DIPEA (2.0 ml, 11 mmol, 1.1 eq) were dissolved in    methylene chloride (15 ml) and cooled in an ice bath.    9-Fluorenylmethyl chloroformate (2.6 g, 10 mmol, 1.0 eq) in    methylene chloride (15 ml) was added, the mixture stirred for 30    minutes then washed with 3N hydrochloric acid (50 ml) twice and    saturated sodium bicarbonate (50 ml). The organics were dried over    magnesium sulfate, and the solvents removed under reduced pressure.    N-Fmoc-2-(decylamino)ethanol (4.6 g, 11 mmol, 108%) was used without    further purification.-   b. N-Fmoc-decylaminoacetaldehyde. To a solution of oxalyl chloride    (12.24 ml) and methylene chloride (50 mL) at −35 to 45° C. was added    DMSO (14.75 g) in methylene chloride (25 mL) over 20 minutes. The    reaction mixture was stirred for 10 minutes at −35 to 45° C. A    solution of N-Fmoc-decylaminoethanol (20.0 g) in methylene chloride    (70 mL) was added over 25 minutes and then stirred 40 minutes at −35    to −45° C. Triethylamine (21.49 g) was then added and the mixture    stirred for 30 minutes at −10 to −20° C. The reaction mixture was    quenched with water (120 mL) followed by concentrated sulfuric acid    (20.0 g) while maintaining the internal temperature at 0-5° C. The    organic layer was isolated and washed with 2% sulfuric acid (100 mL)    followed by water (2×100 mL). The organic solution was distilled    under vacuum at 60° C. to about 100 mL. Heptane (100 mL) was added,    the temperature of the oil bath raised to 80° C. and the    distillation was continued until the residual volume was 100 mL.    More heptane (100 mL) was added and the distillation repeated to a    volume of 100 mL. The heating bath was replaced with a cold water    bath at 15° C. The bath was cooled slowly to 5° C. over 20 minutes    to start the precipitation of the product. The slurry was then    cooled to −5 to −10° C. and the slurry was stirred for 2 hours. The    solid was then collected on a Buchner funnel and washed with cold    (−5° C.) heptane (2×15 mL). The wet solid was dried in vacuo to    yield the aldehyde.-   c. N^(van)—(N-Fmoc-2-n-decylaminoethyl)vancomycin trifluoroacetate.    Vancomycin hydrochloride (12 g, 7.7 mmol, 1.0 eq),    N-Fmoc-2-(n-decylamino)-acetaldehyde (3.2 g, 7.6 mmol, 1.0 eq) and    DIPEA (2.6 ml, 14.9 mmol, 2.0 eq) were stirred at room temperature    in DMF (120 ml) for 90 minutes. Sodium cyanoborohydride (1.4 g, 22    mmol, 3.0 eq) was added, followed by methanol (120 ml) then    trifluoroacetic acid (1.8 ml, 23 mmol, 3.0 eq). The mixture was    stirred for 60 minutes at room temperature, then the methanol    removed under reduced pressure. The resulting solution was added to    600 ml diethyl ether giving a precipitate which was filtered, washed    with ether, and dried under vacuum. The crude product was purified    on a reverse-phase flash column, eluting with 10, 20, 30%    acetonitrile in water (containing 0.1% trifluoroacetic acid) to    remove polar impurities (such as residual vancomycin) then the    product was eluted with 70% acetonitrile in water (containing 0.1%    trifluoroacetic acid) to give 9 g of    N—(N-Fmoc-2-n-decylaminoethyl)vancomycin as its trifluoroacetate    salt (4.3 mmol, 56%).-   d. N^(van)-2-(n-Decylamino)ethyl vancomycin trifluoroacetate.    N—(N-Fmoc-2-n-decylaminoethyl)vancomycin (100 mg) was dissolved in 1    ml DMF (1 ml) and treated with piperidine (200 uL) for 30 minutes.    The mixture was precipitated into ether, centrifuged and washed with    acetonitrile. Reverse-phase preparative HPLC (10-70% acetonitrile in    water containing 0.1% trifluoroacetic acid over 120 minutes) gave    N^(van)-2-(n-decylamino)ethyl vancomycin as its TFA salt.

The intermediate quinuclidine salt of N^(VAN)-decylaminoethyl vancomycinwas prepared as follows.

-   e. N^(van)—(N′-Fmoc-decylaminoethyl)vancomycin. To a 2 L flask    equipped with a mechanical stirrer was added vancomycin    hydrochloride (50.0 g), N-Fmoc-decylaminoacetaldehyde (13.5 g), DMF    (400 mL) and N,N-diisopropylethylamine (11.7 mL). The suspension was    stirred at room temperature for 2 hours, at which time the solids    had dissolved. Methanol (190 mL) followed by trifluoroacetic acid    (10.4 mL) was added. After the reaction mixture had stirred for 5    minutes, borane-pyridine complex (3.33 g) was added in one portion,    and rinsed in with methanol (10 mL). After stirring 4 hours, the    reaction was cooled to 5-10° C. with an ice bath and water (675 mL)    was added at a rate to keep the temperature below 20° C. The    reaction mixture was warmed to room temperature and 10% NaOH was    added to pH 4.2-4.3 (approx 15 mL). The resultant slurry was cooled    in an ice bath for 1 hour, and then the product is collected by    vacuum filtration and washed with cold water (2×100 mL). The wet    solid was dried in vacuo at 50° C. to give the title compound as an    off-white to pale-pink solid.-   f. N^(VAN)-(decylaminoethyl)vancomycin quinuclidine salt.    N^(van)—(N′-Fmoc-decylaminoethyl)vancomycin (88 g, 42 mmol) was    dissolved in DMF (500 mL) by stirring at room temperature for 1    hour. Quinuclidine (9.4 g, 84 mmol) was added, and the reaction    mixture stirred for 18 hours. The DMF was removed in vacuo and the    solid was triturated with acetonitrile (700 mL) for 3 hours. The    solid was collected on a Buchner funnel and triturated with    acetonitrile (200 mL) for 16 hours. More acetonitrile (700 mL) was    added at this time, and the solid was collected on a Buchner funnel,    washed with acetonitrile (500 mL), and then resuspended in    acetonitrile (500 mL). After stirring for 2 hours, the solid was    collected on a Buchner funnel and dried in vacuo to give the title    compound.

Example 3 Preparation of Compound 12 Formula II wherein R³ is —OH; R⁵N-(phosphonomethyl)-aminomethyl; R¹⁹ is hydrogen, and R²⁰ is—CH₂CH₂—S—(CH₂)₉CH₃

(Aminomethyl)phosphonic acid (0.295 g, 266 mmol) anddiisopropylethylamine (0.649 ml, 3.72 mmol) were combined in water (5ml) and stirred until homogeneous. Formaldehyde (37% solution in H₂O;0.044 ml, 0.585 mmol) and acetonitrile (5 ml) were then added to thereaction mixture. After approximately 15 minutes bothN^(VAN)-(2-decylthio)ethyl vancomycin bistrifluoroacetate (1 g, 0.53mmol) and diisopropylethylamine (0.649 ml, 3.72 mmol) were added to thereaction mixture. The reaction was stirred at room temperature forapproximately 18 hrs, at which time the reaction mixture was dilutedwith ACN (40 ml) and centrifuged. The supernatant was discarded and theremaining pellet containing desired product was dissolved in 50% aqueousacetonitrile (10 ml) and purified by reverse phase preparative HPLC togive the title compound. MS calculated (M+) 1772.7; found (MH+) 1773.4.

Using the above procedures and the appropriate starting materials thecompounds shown in Table I were prepared. The mass spectral data forthese compounds were as follows:

Compound No. MW (freebase) Observed MH⁺ 1 1725.63 1726.6 2 1726.621727.5 3 1742.68 1743.6 4 1724.64 1725.6 5 1742.96 1743.6 6 1786.031786.4 7 1785.04 1785.8 8 1799.07 1799.7 9 1770.74 1771.8 10 1772.991774.3 11 1755.66 1756.6 12 1772.71 1773.4 13 1756.64 1757.6 14 1754.671755.7 15 1772.99 1773.7 16 1816.06 1816.5 17 1815.01 1816.2 18 1829.101829.8 19 1878.1 1878.2 20 1802.74 1803.5 21 1830.75 1831.7 22 1849.661850.6 23 1800.76 1801.6 24 1801.04 1801.6 25 1932.86 1934.0 26 1880.121880.7

Example 4 Preparation of an Intermediate Useful for Preparing a Compoundof the Invention Formula II wherein R^(c) is —OH; R³ is H; R¹⁹ isHydrogen, and R²⁰ is 4-(4-chlorophenyl)benzyl

A three liter 3-necked flask was fitted with a condenser, nitrogen inletand overhead mechanical stirring apparatus. The flask was charged withpulverized A82846B acetate salt (20.0 g, 1.21×10⁻⁵ mol) and methanol(1000 ml) under a nitrogen atmosphere, 4′-chlorobiphenylcarboxaldehyde(2.88 g, 1.33×10⁻² mol, 1.1 eq.) was added to this stirred mixture,followed by methanol (500 ml). Finally, sodium cyanoborohydride (0.84 g,1.33×10⁻² mol, 1.1 eq.) was added followed by methanol (500 ml). Theresulting mixture was heated to reflux (about 65° C.).

After 1 hour at reflux, the reaction mixture attained homogeneity. After25 hours at reflux, the heat source was removed and the clear reactionmixture was measured with a pH meter (6.97 at 58.0° C.). 1N NaOH (22.8ml) was added dropwise to adjust the pH to 9.0 (at 54.7° C.). The flaskwas equipped with a distillation head and the mixture was concentratedunder partial vacuum to a weight of 322.3 grams while maintaining thepot temperature between 40°-45° C.

The distillation head was replaced with an addition funnel containing500 ml of isopropanol (IPA). The IPA was added dropwise to the roomtemperature solution over 1 hour. After approximately ⅓ of the IPA wasadded, a granular precipitate formed. The remaining IPA was added at afaster rate after precipitation had commenced. The flask was weighed andfound to hold 714.4 grams of the IPA/methanol slurry.

The flask was re-equipped with a still-head and distilled under partialvacuum to remove the remaining methanol. The resulting slurry (377.8 g)was allowed to chill in the freezer overnight. The crude product wasfiltered through a polypropylene pad and rinsed twice with 25 ml of coldIPA. After pulling dry on the funnel for 5 minutes, the material wasplaced in the vacuum oven to dry to 40° C. A light pink solid (22.87 g(theory=22.43 g)) was recovered. HPLC analysis versus a standardindicated 68.0% weight percent of the title compound(4-[4-chlorophenyl]benzyl-A82846B] in the crude solid, which translatedinto a corrected crude yield of 69.3%.

The products of the reaction were analyzed by reverse-phase HPLCutilizing a Zorbax SB-C₁₈ column with ultra-violet light (UV; 230 nm)detection. A 20 minute gradient solvent system consisting of 95% aqueousbuffer/5% CH₃CN at time=0 minutes to 40% aqueous buffer/60% CH₃CN attime=30 minutes was used, where the aqueous buffer was TEAP (5 ml CH₃CN,3 ml phosphoric acid in 1000 ml water).

The intermediate A82846B acetate salt can be prepared as described inU.S. Pat. No. 5,840,684.

Using procedures described hereinabove, the product of Example 4 can beconverted to a compound of the invention wherein R³ and/or R⁵ is asubstituent that comprises one or more phosphono groups.

Example 5 Determination of Antibacterial Activity

A. In Vitro Determination of Antibacterial Activity

1. Determination of Minimal Inhibitory Concentrations (MICs)

Bacterial strains were obtained from either American Type Tissue CultureCollection (ATCC), Stanford University Hospital (SU), Kaiser PermanenteRegional Laboratory in Berkeley (KPB), Mass. General Hospital (GH), theCenters for Disease Control (CDC), the San Francisco Veterans'Administration Hospital (SFVA) or the University of California SanFrancisco Hospital (UCSF). Vancomycin resistant enterococci werephenotyped as Van A or Van B based on their sensitivity to teicoplanin.Some vancomycin resistant enterococci that had been genotyped as Van A,Van B, Van C1 or Van C2 were obtained from the Mayo Clinic.

Minimal inhibitory concentrations (MICs) were measured in amicrodilution broth procedure under NCCLS guidelines. Routinely, thecompounds were serially diluted into Mueller-Hinton broth in 96-wellmicrotiter plates. Overnight cultures of bacterial strains were dilutedbased on absorbance at 600 nm so that the final concentration in eachwell was 5×10⁵ cfu/ml. Plates were returned to a 35° C. incubator. Thefollowing day (or 24 hours in the case of Enterococci strains), MICswere determined by visual inspection of the plates. Strains routinelytested in the initial screen included methicillin-sensitiveStaphylococcus aureus (MSSA), methicillin-resistant Staphylococcusaureus, methicillin-sensitive Staphylococcus epidermidis (MSSE),methicillin-resistant Staphylococcus epidermidis (MSSE), vancomycinsensitive Enterococcus faecium (VSE Fm), vancomycin sensitiveEnterococcus faecalis (VSE Fs), vancomycin resistant Enterococcusfaecium also resistant to teicoplanin (VRE Fm Van A), vancomycinresistant Enterococcus faecium sensitive to teicoplanin (VRE Fm Van B),vancomycin resistant Enterococcus faecalis also resistant to teicoplanin(VRE Fs Van A), vancomycin resistant Enterococcus faecalis sensitive toteicoplanin (VRE Fs Van B), enterococcus gallinarium of the Van Agenotype (VRE Gm Van A), enterococcus gallinarium of the Van C-1genotype (VRE Gm Van C-1), enterococcus casseliflavus of the Van C-2genotype (VRE Cs Van C-2), enterococcus flavescens of the Van C-2genotype (VRE Fv Van C-2), and penicillin-sensitive Streptococcuspneumoniae (PSSP) and penicillin-resistant Streptococcus pneumoniae(PSRP). Because of the inability of PSSP and PSRP to grow well inMueller-Hinton broth, MICs with those strains were determined usingeither TSA broth supplemented with defibrinated blood or blood agarplates. Compounds which had significant activity against the strainsmentioned above were then tested for MIC values in a larger panel ofclinical isolates including the species listed above as well asnon-speciated coagulase negative Staphylococcus both sensitive andresistant to methicillin (MS-CNS and MR-CNS). In addition, they weretested for MICs against gram negative organisms, such as Escherichiacoli and Pseudomonas aeruginosa.

2. Determination of Kill Time

Experiments to determine the time required to kill the bacteria wereconducted as described in Lorian, “Antibiotics in Laboratory Medicine”,Fourth Edition, Williams and Wilkins (1991). These experiments wereconducted normally with both staphylococcus and enterococcus strains.

Briefly, several colonies were selected from an agar plate and grown at35° C. under constant agitation until it achieved a turbidity ofapproximately 1.5 and 10⁸ CFU/ml. The sample was then diluted to about6×10⁶ CFU/ml and incubated at 35° C. under constant agitation wascontinued. At various times aliquots were removed and five ten-foldserial dilutions were performed. The pour plate method was used todetermine the number of colony forming units (CFUs).

In general, the compounds of the invention were active in the abovetests in vitro tests and demonstrated a broad spectrum of activity.

B. In Vivo Determination of Antibacterial Activity

1. Acute Tolerability Studies in Mice

In these studies, a compound of this invention was administered eitherintravenously or subcutaneously and observed for 5-15 minutes. If therewere no adverse effects, the dose was increased in a second group ofmice. This dose incrementation continued until mortality occurred, orthe dose was maximized. Generally, dosing began at 20 mg/kg andincreased by 20 mg/kg each time until the maximum tolerated dose (MTD)is achieved.

2. Bioavailability Studies in Mice

Mice were administered a compound of this invention either intravenouslyor subcutaneously at a therapeutic dose (in general, approximately 50mg/kg). Groups of animals were placed in metabolic cages so that urineand feces could be collected for analysis. Groups of animals (n=3) weresacrificed at various times (10 min, 1 hour and 4 hours). Blood wascollected by cardiac puncture and the following organs wereharvested-lung, liver, heart, brain, kidney, and spleen. Tissues wereweighed and prepared for HPLC analysis. HPLC analysis on the tissuehomogenates and fluids was used to determine the concentration of thetest compound or lit present. Metabolic products resulting from changesto the test compound were also determined at this juncture.

3. Mouse Septicemia Model

In this model, an appropriately virulent strain of bacteria (mostcommonly S. aureus, or E. Faecalis or E. Faecium) was administered tomice (N=5 to 10 mice per group) intraperitoneally. The bacteria wascombined with hog gastric mucin to enhance virulence. The dose ofbacteria (normally 10⁵-10⁷) was that sufficient to induce mortality inall of the mice over a three day period. One hour after the bacteria wasadministered, a compound of this invention was administered in a singledose either IV or subcutaneously. Each dose was administered to groupsof 5 to 10 mice, at doses that typically ranged from a maximum of about20 mg/kg to a minimum of less than 1 mg/kg. A positive control (normallyvancomycin with vancomycin sensitive strains) was administered in eachexperiment. The dose at which approximately 50% of the animals are savedwas calculated from the results.

4. Neutropenic Thigh Model

In this model, antibacterial activity of a compound of this inventionwas evaluated against an appropriately virulent strain of bacteria (mostcommonly S. aureus, or E. Faecalis or E. Faecium, sensitive or resistantto vancomycin). Mice were initially rendered neutropenic byadministration of cyclophosphamide at 200 mg/kg on day 0 and day 2. Onday 4 they were infected in the left anterior thigh by an IM injectionof a single dose of bacteria. The mice were then administered the testcompound one hour after the bacteria and at various later times(normally 1, 2.5, 4 and 24 hours) the mice were sacrificed (3 per timepoint) and the thigh excised, homogenized and the number of CFUs (colonyforming units) were determined by plating. Blood was also plated todetermine the CFUs in the blood.

5. Pharmacokinetic Studies

The rate at which a compound of this invention is removed from the bloodcan be determined in either rats or mice. In rats, the test animals werecannulated in the jugular vein. The test compound was administered viatail vein injection, and at various time points (normally 5, 15, 30, 60minutes and 2, 4, 6 and 24 hours) blood was withdrawn from the cannulaIn mice, the test compound was also administered via tail veininjection, and at various time points. Blood was normally obtained bycardiac puncture. The concentration of the remaining test compound wasdetermined by HPLC.

In general, the compounds of the invention were active in the above testin vivo and demonstrated a broad spectrum of activity.

Example 6

Determination of Tissue Accumulation

A. Tissue Distribution Using Radiolabeled Compound

This procedure is used to examine the tissue distribution, excretion andmetabolism of a radiolabeled test compound in both male and female ratsfollowing intravenous infusion at 10 mg/kg. Male and femaleSprague-Dawley rats (n=2 per sex per compound) are dosed with ³H-labeledtest compound at 10 (400 μCi/kg) and 12.5 mg/kg (100 μCi/kg),respectively, via intravenous infusion (˜2 min). The test compound isformulated in 5% hydroxypropyl-β-cyclodextrin as 2.5 mg/ml solution.Urine and feces are cage collected over 24 hours period. At 24 hoursafter dosing, animals are sacrificed and tissues are removed. Serum,urine and tissues are analyzed for total radioactivity by oxidationfollowed by liquid scintillation counting. Urine and selected tissuessamples are extracted and analyzed by reverse phase HPLC withradioactive flow detector for the presence of potential metabolites.

B. Tissue Accumulation Following Single Dose

This procedure is used to evaluate tissue distribution of a testcompound in rats following single dose administration by infusion. MaleSprague-Dawley rats (n=3 per dose groups) are dosed with 50 mg/kg of atest compound. Two formulations are used: 30% PEG 400 and 10%sulfobutylether-β-cyclodextrin. Urine samples are cage collected over 24hours. Blood samples are collected for serum chemistry and concentrationdetermination. Liver and kidneys are removed for histology evaluation.One kidney and part of the liver are homogenized for concentrationanalysis using reverse phase HPLC with UV detection. Drug concentrationsin urine and serum samples are determined by LC-MS analysis.

C. Tissue Distribution Following Multiple Doses

This procedure is used to evaluate the potential tissue accumulation ofa test compound in rats following multiple dose administration byintravenous infusion. Male and female Sprague-Dawley rats (n=4 per sexper dose group) are dosed with a test compound at 12.5, 25 and 50 mg/kgper day for seven days. Animals are sacrificed at day 1 (n=3 per sex perdose group) following the last dose administered. One animal per sex perdose group is retained as recovery animal and sacrificed at day 7following the last dose administered. The test compound is formulated in5% hydroxypropyl-β-cyclodextrin or 1% sucrose/4.5% dextrose. Urinesamples are cage collected at days 1 and 7 post-dose. Blood samples arecollected for serum chemistry and concentration determination. Liver andkidneys are removed for histology evaluation. One kidney and part of theliver are homogenized for concentration analysis using reverse phaseHPLC with UV detection. Drug concentrations in urine and serum samplesare determined by LC-MS analysis.

While the present invention has been described with reference to thespecific embodiments thereof, it should be understood by those skilledin the art that various changes may be made and equivalents may besubstituted without departing from the true spirit and scope of theinvention. In addition, many modifications may be made to adapt aparticular situation, material, composition of matter, process, processstep or steps, to the objective, spirit and scope of the presentinvention. All such modifications are intended to be within the scope ofthe claims appended hereto. Additionally, all publications, patents, andpatent documents cited hereinabove are incorporated by reference hereinin full, as though individually incorporated by reference.

1. A method for treating a bacterial infection in a human, the methodcomprising administering to the human a compound of the formula:

or a pharmaceutically acceptable salt thereof.
 2. The method of claim 1,wherein the compound is a hydrochloride salt.
 3. The method of claim 1,wherein the compound or a pharmaceutically acceptable salt thereof isadministered intravenously.
 4. The method of claim 1, wherein thecompound or a pharmaceutically acceptable salt thereof is administeredonce per day.
 5. A method for treating a staphylococcal infection in ahuman, the method comprising administering to the human a compound ofthe formula:

or a pharmaceutically acceptable salt thereof.
 6. The method of claim 5,wherein the compound is a hydrochloride salt.
 7. The method of claim 5,wherein the compound or a pharmaceutically acceptable salt thereof isadministered intravenously.
 8. The method of claim 5, wherein thecompound or a pharmaceutically acceptable salt thereof is administeredonce per day.
 9. The method of claim 5, wherein the staphylococcalinfection is caused by Staphylococcus aureus.
 10. The method of claim 5,wherein the staphylococcal infection is caused by methicillin-resistantStaphylococcus aureus.